LOCUS       DR274_loxP_hygR_        4621 bp    DNA        circular     02-AUG-2023
DEFINITION  .
ACCESSION   
VERSION     
SOURCE      .
  ORGANISM  .
COMMENT     PCR reaction:
COMMENT     template: pDD363 SEC loxP [3432]
COMMENT     New Primer tGGTCTCaGGTCcATTTTTGCTTTCGTCGTAAATC Tm1: 52.9°C Tm2:
            65.5°C.
COMMENT     New Primer tggtctcagaccttagaggtcctcctcgg Tm1: 56.3°C Tm2: 66.9°C.
COMMENT     Product length: 4621
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     misc_feature    4512..4518
                     /locus_tag="SapI"
                     /label="SapI"
                     /ApEinfo_label="SapI"
                     /ApEinfo_fwdcolor="#ea17ff"
                     /ApEinfo_revcolor="#ea17ff"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(4593..4621)
                     /PCR_conditions="primer
                     sequence:tggtctcagaccttagaggtcctcctcgg"
                     /locus_tag="New Primer"
                     /label="New Primer"
                     /ApEinfo_label="New Primer"
                     /ApEinfo_fwdcolor="#408000"
                     /ApEinfo_revcolor="#800080"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     1..35
                     /PCR_conditions="primer
                     sequence:tGGTCTCaGGTCcATTTTTGCTTTCGTCGTAAATC"
                     /locus_tag="New Primer(1)"
                     /label="New Primer(1)"
                     /ApEinfo_label="New Primer"
                     /ApEinfo_fwdcolor="#408000"
                     /ApEinfo_revcolor="#800080"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     4300..4317
                     /locus_tag="M13-fwd"
                     /label="M13-fwd"
                     /ApEinfo_label="M13-fwd"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    1866..1872
                     /locus_tag="SapI(1)"
                     /label="SapI(1)"
                     /ApEinfo_label="SapI"
                     /ApEinfo_fwdcolor="#ea17ff"
                     /ApEinfo_revcolor="#ea17ff"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(2025..2045)
                     /locus_tag="M13-rev"
                     /label="M13-rev"
                     /ApEinfo_label="M13-rev"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(2005..2026)
                     /locus_tag="T7"
                     /label="T7"
                     /ApEinfo_label="T7"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             2158..2964
                     /locus_tag="KanR"
                     /label="KanR"
                     /ApEinfo_label="KanR"
                     /ApEinfo_fwdcolor="yellow"
                     /ApEinfo_revcolor="yellow"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     intron          4523..4533
                     /locus_tag="intron"
                     /label="intron"
                     /ApEinfo_label="intron"
                     /ApEinfo_fwdcolor="#cc66ff"
                     /ApEinfo_revcolor="#ff00ff"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    4520..4522
                     /locus_tag="AAG overlap"
                     /label="AAG overlap"
                     /ApEinfo_label="AAG overlap"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_recomb     4355..4497
                     /locus_tag="attP2r"
                     /label="attP2r"
                     /ApEinfo_label="attP2r"
                     /ApEinfo_fwdcolor="#ffcccc"
                     /ApEinfo_revcolor="#cccccc"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    4577..4606
                     /locus_tag="myc"
                     /label="myc"
                     /ApEinfo_label="myc"
                     /ApEinfo_fwdcolor="#6666ff"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_recomb     1880..1968
                     /locus_tag="attP3"
                     /label="attP3"
                     /ApEinfo_label="attP3"
                     /ApEinfo_fwdcolor="#ffcccc"
                     /ApEinfo_revcolor="#cccccc"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             419..1444
                     /note="hygromycinR"
                     /locus_tag="hygR"
                     /label="hygR"
                     /ApEinfo_label="hygR"
                     /ApEinfo_fwdcolor="#006666"
                     /ApEinfo_revcolor="#006666"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     intron          4568..4573
                     /locus_tag="intron(1)"
                     /label="intron(1)"
                     /ApEinfo_label="intron"
                     /ApEinfo_fwdcolor="#cc66ff"
                     /ApEinfo_revcolor="#ff00ff"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    4534..4567
                     /locus_tag="loxP"
                     /label="loxP"
                     /ApEinfo_label="loxP"
                     /ApEinfo_fwdcolor="#ff8000"
                     /ApEinfo_revcolor="#ff8000"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    1822..1855
                     /locus_tag="loxP(1)"
                     /label="loxP(1)"
                     /ApEinfo_label="loxP"
                     /ApEinfo_fwdcolor="#ff8000"
                     /ApEinfo_revcolor="#ff8000"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     promoter        complement(13..13)
                     /locus_tag="hsp16-41 promoter"
                     /label="hsp16-41 promoter"
                     /ApEinfo_label="hsp16-41 promoter"
                     /ApEinfo_fwdcolor="#0080c0"
                     /ApEinfo_revcolor="#0080c0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     intron          1856..1861
                     /locus_tag="intron(2)"
                     /label="intron(2)"
                     /ApEinfo_label="intron"
                     /ApEinfo_fwdcolor="#cc66ff"
                     /ApEinfo_revcolor="#ff00ff"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     terminator      4610..4611
                     /locus_tag="let-858 Terminator"
                     /label="let-858 Terminator"
                     /ApEinfo_label="let-858 Terminator"
                     /ApEinfo_fwdcolor="#cc0000"
                     /ApEinfo_revcolor="#cc0000"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     3'UTR           1470..1821
                     /locus_tag="unc-54 3'UTR"
                     /label="unc-54 3'UTR"
                     /ApEinfo_label="unc-54 3'UTR"
                     /ApEinfo_fwdcolor="#990000"
                     /ApEinfo_revcolor="#cc0000"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    4574..4576
                     /locus_tag="spacer"
                     /label="spacer"
                     /ApEinfo_label="spacer"
                     /ApEinfo_fwdcolor="#cccccc"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     promoter        14..376
                     /note="Prps-0"
                     /note="/ApEinfo_fwdcolor=pink"
                     /note=""/ApEinfo_revcolor=pink; /ApEinfo_fwdcolor=pink;"
                     /locus_tag="Prps-0"
                     /label="Prps-0"
                     /ApEinfo_label="Prps-0"
                     /ApEinfo_fwdcolor="#339933"
                     /ApEinfo_revcolor="#339933"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    1862..1864
                     /locus_tag="GGT overlap"
                     /label="GGT overlap"
                     /ApEinfo_label="GGT overlap"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_recomb     4498..4511
                     /locus_tag="attB2r"
                     /label="attB2r"
                     /ApEinfo_label="attB2r"
                     /ApEinfo_fwdcolor="#cccccc"
                     /ApEinfo_revcolor="#cccccc"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_recomb     1873..1879
                     /locus_tag="attB3"
                     /label="attB3"
                     /ApEinfo_label="attB3"
                     /ApEinfo_fwdcolor="#cccccc"
                     /ApEinfo_revcolor="#cccccc"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
ORIGIN
        1 tGGTCTCaGG TCcATTTTTG CTTTCGTCGT AAATCTACAC ACGCGTCTCT TCCGTGCGAG
       61 AGTCCAAGCC AGCAGCCAAA TTCGTTGACT GAGTATTCAA CGTTTATACG TTGTCGGCAA
      121 CGAGAAATAG GAAAATGCAT CGGGAAATGT TCTTTTTTCG ATTTTTTCCA AGGTTTTGAC
      181 AAATTTTACC ACGAATTTTG CTATGTTTTC AATTAAAAAA TATGTTATTC AACTGTTTCT
      241 ATGAGGAAAA TAAGGCTTTG CATGTAATTT TCTTATTCAG CATAATTTTT AATTAATTTG
      301 AATTTTCTGT CCTAACGTTT ATTTTGTTTT CTTGGTTATG ACTGATCTGA AATTAATTTT
      361 TGAATTTTAA GGTAATATGT CAGGCGGTGC CGCAAGTTTG TACAAAAAAG CAGGCTCCAT
      421 GAAAAAGCCT GAACTCACCG CGACGTCTGT CGAGAAGTTT CTGATCGAAA AGTTCGACAG
      481 CGTCTCCGAC CTGATGCAGC TCTCGGAGGG CGAAGAATCT CGTGCTTTCA GCTTCGATGT
      541 AGGAGGGCGT GGATATGTCC TGCGGGTAAA TAGCTGCGCC GATGGTTTCT ACAAAGATCG
      601 TTATGTTTAT CGGCACTTTG CATCGGCCGC GCTCCCGATT CCGGAAGTGC TTGACATTGG
      661 GGAATTCAGC GAGAGCCTGA CCTATTGCAT CTCCCGCCGT GCACAGGGTG TCACGTTGCA
      721 AGACCTGCCT GAAACCGAAC TGCCCGCTGT TCTGCAGCCG GTCGCGGAGG CCATGGATGC
      781 GATCGCTGCG GCCGATCTTA GCCAGACGAG CGGGTTCGGC CCATTCGGAC CGCAAGGAAT
      841 CGGTCAATAC ACTACATGGC GTGATTTCAT ATGCGCGATT GCTGATCCCC ATGTGTATCA
      901 CTGGCAAACT GTGATGGACG ACACCGTCAG TGCGTCCGTC GCGCAGGCTC TCGATGAGCT
      961 GATGCTTTGG GCCGAGGACT GCCCCGAAGT CCGGCACCTC GTGCACGCGG ATTTCGGCTC
     1021 CAACAATGTC CTGACGGACA ATGGCCGCAT AACAGCGGTC ATTGACTGGA GCGAGGCGAT
     1081 GTTCGGGGAT TCCCAATACG AGGTCGCCAA CATCTTCTTC TGGAGGCCGT GGTTGGCTTG
     1141 TATGGAGCAG CAGACGCGCT ACTTCGAGCG GAGGCATCCG GAGCTTGCAG GATCGCCGCG
     1201 GCTCCGGGCG TATATGCTCC GCATTGGTCT TGACCAACTC TATCAGAGCT TGGTTGACGG
     1261 CAATTTCGAT GATGCAGCTT GGGCGCAGGG TCGATGCGAC GCAATCGTCC GATCCGGAGC
     1321 CGGGACTGTC GGGCGTACAC AAATCGCCCG CAGAAGCGCG GCCGTCTGGA CCGATGGCTG
     1381 TGTAGAAGTA CTCGCCGATA GTGGAAACCG ACGCCCCAGC ACTCGTCCGA GGGCAAAGGA
     1441 ATAGACCCAG CTTTCTTGTA CAAAGTGGGT CCAATTACTC TTCAACATCC CTACATGCTC
     1501 TTTCTCCCTG TGCTCCCACC CCCTATTTTT GTTATTATCA AAAAACTTCT CTTAATTTCT
     1561 TTGTTTTTTA GCTTCTTTTA AGTCACCTCT AACAATGAAA TTGTGTAGAT TCAAAAATAG
     1621 AATTAATTCG TAATAAAAAG TCGAAAAAAA TTGTGCTCCC TCCCCCCATT AATAATAATT
     1681 CTATCCCAAA ATCTACACAA TGTTCTGTGT ACACTTCTTA TGTTTTTTAC TTCTGATAAA
     1741 TTTTTTTGAA ACATCATAGA AAAAACCGCA CACAAAATAC CTTATCATAT GTTACGTTTC
     1801 AGTTTATGAC CGCAATTTTT AATAACTTCG TATAGCATAC ATTATACGAA GTTATtttca
     1861 gGGTcGAAGA GCCAACTTTA TTATACAAAG TTGGCATTAT AAAAAAGCAT TGCTTATCAA
     1921 TTTGTTGCAA CGAACAGGTC ACTATCAGTC AAAATAAAAT CATTATTTGG AGCTCCATGG
     1981 TAGCGTTAAC GCGGCCGCGA TATCCCCTAT AGTGAGTCGT ATTACATGGT CATAGCTGTT
     2041 TCCTGGCAGC TCTGGCCCGT GTCTCAAAAT CTCTGATGTT ACATTGCACA AGATAAAAAT
     2101 ATATCATCAT GAACAATAAA ACTGTCTGCT TACATAAACA GTAATACAAG GGGTGTTATG
     2161 AGCCATATTC AACGGGAAAC GTCGAGGCCG CGATTAAATT CCAACATGGA TGCTGATTTA
     2221 TATGGGTATA AATGGGCTCG CGATAATGTC GGGCAATCAG GTGCGACAAT CTATCGCTTG
     2281 TATGGGAAGC CCGATGCGCC AGAGTTGTTT CTGAAACATG GCAAAGGTAG CGTTGCCAAT
     2341 GATGTTACAG ATGAGATGGT CAGACTAAAC TGGCTGACGG AATTTATGCC TCTTCCGACC
     2401 ATCAAGCATT TTATCCGTAC TCCTGATGAT GCATGGTTAC TCACCACTGC GATCCCCGGA
     2461 AAAACAGCAT TCCAGGTATT AGAAGAATAT CCTGATTCAG GTGAAAATAT TGTTGATGCG
     2521 CTGGCAGTGT TCCTGCGCCG GTTGCATTCG ATTCCTGTTT GTAATTGTCC TTTTAACAGC
     2581 GATCGCGTAT TTCGTCTCGC TCAGGCGCAA TCACGAATGA ATAACGGTTT GGTTGATGCG
     2641 AGTGATTTTG ATGACGAGCG TAATGGCTGG CCTGTTGAAC AAGTCTGGAA AGAAATGCAT
     2701 AAACTTTTGC CATTCTCACC GGATTCAGTC GTCACTCATG GTGATTTCTC ACTTGATAAC
     2761 CTTATTTTTG ACGAGGGGAA ATTAATAGGT TGTATTGATG TTGGACGAGT CGGAATCGCA
     2821 GACCGATACC AGGATCTTGC CATCCTATGG AACTGCCTCG GTGAGTTTTC TCCTTCATTA
     2881 CAGAAACGGC TTTTTCAAAA ATATGGTATT GATAATCCTG ATATGAATAA ATTGCAGTTT
     2941 CATTTGATGC TCGATGAGTT TTTCTAATCA GAATTGGTTA ATTGGTTGTA ACACTGGCAG
     3001 AGCATTACGC TGACTTGACG GGACGGCGCA AGCTCATGAC CAAAATCCCT TAACGTGAGT
     3061 TACGCGTCGT TCCACTGAGC GTCAGACCCC GTAGAAAAGA TCAAAGGATC TTCTTGAGAT
     3121 CCTTTTTTTC TGCGCGTAAT CTGCTGCTTG CAAACAAAAA AACCACCGCT ACCAGCGGTG
     3181 GTTTGTTTGC CGGATCAAGA GCTACCAACT CTTTTTCCGA AGGTAACTGG CTTCAGCAGA
     3241 GCGCAGATAC CAAATACTGT CCTTCTAGTG TAGCCGTAGT TAGGCCACCA CTTCAAGAAC
     3301 TCTGTAGCAC CGCCTACATA CCTCGCTCTG CTAATCCTGT TACCAGTGGC TGCTGCCAGT
     3361 GGCGATAAGT CGTGTCTTAC CGGGTTGGAC TCAAGACGAT AGTTACCGGA TAAGGCGCAG
     3421 CGGTCGGGCT GAACGGGGGG TTCGTGCACA CAGCCCAGCT TGGAGCGAAC GACCTACACC
     3481 GAACTGAGAT ACCTACAGCG TGAGCATTGA GAAAGCGCCA CGCTTCCCGA AGGGAGAAAG
     3541 GCGGACAGGT ATCCGGTAAG CGGCAGGGTC GGAACAGGAG AGCGCACGAG GGAGCTTCCA
     3601 GGGGGAAACG CCTGGTATCT TTATAGTCCT GTCGGGTTTC GCCACCTCTG ACTTGAGCGT
     3661 CGATTTTTGT GATGCTCGTC AGGGGGGCGG AGCCTATGGA AAAACGCCAG CAACGCGGCC
     3721 TTTTTACGGT TCCTGGCCTT TTGCTGGCCT TTTGCTCACA TGTTCTTTCC TGCGTTATCC
     3781 CCTGATTCTG TGGATAACCG TATTACCGCC TTTGAGTGAG CTGATACCGC TCGCCGCAGC
     3841 CGAACGACCG AGCGCAGCGA GTCAGTGAGC GAGGAAGCGG AAGAGCGCCC AATACGCAAA
     3901 CCGCCTCTCC CCGCGCGTTG GCCGATTCAT TAATGCAGCT GGCACGACAG GTTTCCCGAC
     3961 TGGAAAGCGG GCAGTGAGCG CAACGCAATT AATACGCGTA CCGCTAGCCA GGAAGAGTTT
     4021 GTAGAAACGC AAAAAGGCCA TCCGTCAGGA TGGCCTTCTG CTTAGTTTGA TGCCTGGCAG
     4081 TTTATGGCGG GCGTCCTGCC CGCCACCCTC CGGGCCGTTG CTTCACAACG TTCAAATCCG
     4141 CTCCCGGCGG ATTTGTCCTA CTCAGGAGAG CGTTCACCGA CAAACAACAG ATAAAACGAA
     4201 AGGCCCAGTC TTCCGACTGA GCCTTTCGTT TTATTTGATG CCTGGCAGTT CCCTACTCTC
     4261 GCGTTAACGC TAGCATGGAT GTTTTCCCAG TCACGACGTT GTAAAACGAC GGCCAGTCTT
     4321 AAGCTCGGGC CCTGCAGCTC TAGAGCTCGA ATTCTACAGG TCACTAATAC CATCTAAGTA
     4381 GTTGATTCAT AGTGACTGCA TATGTTGTGT TTTACAGTAT TATGTAGTCT GTTTTTTATG
     4441 CAAAATCTAA TTTAATATAT TGATATTTAT ATCATTTTAC GTTTCTCGTT CAACTTTCTT
     4501 GTACAAAGTG GGCTCTTCgA AGgtaagttt aaaATAACTT CGTATAGCAT ACATTATACG
     4561 AAGTTATttt cagGGAGAGC AAAAGCTTAT CTCCGAGGAG GACCTCTAAG Gtctgagacc
     4621 a     
//
