LOCUS       Exported                4540 bp ds-DNA     circular SYN 24-FEB-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4540)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Mar 3, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4540
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..88
                     /locus_tag="misc_feature"
                     /label=misc_feature
     misc_feature    7..23
                     /locus_tag="-43 AT rich region"
                     /label=-43 AT rich region
                     /note="-43 AT rich region"
     misc_feature    25..30
                     /locus_tag="-35 box"
                     /label=-35 box
                     /note="-35 box"
     misc_feature    30..48
                     /locus_tag="lacO(1)"
                     /label=lacO
                     /label=lacO(1)
                     /note="lacO"
     misc_feature    48..53
                     /locus_tag="-10 box"
                     /label=-10 box
                     /note="-10 box"
     misc_feature    60
                     /locus_tag="transcription start"
                     /label=transcription start
                     /note="transcription start"
     misc_feature    62..80
                     /locus_tag="lacO"
                     /label=lacO
                     /note="lacO"
     misc_feature    89..113
                     /locus_tag="misc_feature(2)"
                     /label=misc_feature(2)
     misc_feature    96..106
                     /locus_tag="RBS"
                     /label=RBS
                     /note="RBS"
     misc_feature    114..192
                     /locus_tag="misc_feature(1)"
                     /label=misc_feature(1)
     misc_feature    115..192
                     /locus_tag="His14-TAG"
                     /label=His14-TAG
                     /note="His14-TAG"
     misc_feature    205..249
                     /locus_tag="Biotin-tag"
                     /label=Biotin-tag
                     /note="Biotin-tag"
     misc_feature    329..384
                     /locus_tag="Spacer 21 aa"
                     /label=Spacer 21 aa
                     /note="Spacer 21 aa"
     misc_feature    385..732
                     /locus_tag="anti-GFP nanobody 3K1K"
                     /label=anti-GFP nanobody 3K1K
                     /note="anti-GFP nanobody 3K1K"
     misc_feature    784..878
                     /locus_tag="lambda t0 trx termination region"
                     /label=lambda t0 trx termination region
                     /note="lambda t0 trx termination region"
     misc_feature    937..1752
                     /locus_tag="KanR"
                     /label=KanR
                     /note="KanR"
     misc_feature    complement(1965..2062)
                     /locus_tag="rrnB T1 trx termination region"
                     /label=rrnB T1 trx termination region
                     /note="rrnB T1 trx termination region"
     primer_bind     2029..2053
                     /label=Primer 1
     misc_feature    complement(2110..3211)
                     /locus_tag="lacI E.coli genomic sequence"
                     /label=lacI E.coli genomic sequence
                     /note="lacI E.coli genomic sequence"
     misc_feature    complement(2151..3110)
                     /locus_tag="lacI"
                     /label=lacI
                     /note="lacI"
     primer_bind     3313..3333
                     /label=Primer 2
     misc_feature    3360..3462
                     /locus_tag="part of H-strand Y effector site"
                     /label=part of H-strand Y effector site
                     /note="part of H-strand Y effector site"
     primer_bind     complement(3784..3809)
                     /label=Primer 3
     primer_bind     3784..3809
                     /label=Primer 4
     misc_feature    3829..4417
                     /locus_tag="ColE1 origin"
                     /label=ColE1 origin
                     /note="ColE1 origin"
     misc_feature    3830
                     /locus_tag="replication origin"
                     /label=replication origin
                     /note="replication origin"
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aaccatgagc
      121 aagcatcacc atcattcagg ccatcaccat accggacacc accatcattc aggcagtcat
      181 caccataccg gcggatcatc cggaggtctg aatgatatct ttgaagcaca aaagattgaa
      241 tggcatgaag gctctggatc tgcaactggt tcaggaggca gtgggggttc cggtacaccg
      301 ggcagttcta cgagtggcac ctctgcagga tccggttctg gtggtagcgg tggttcagga
      361 ggcagtgggg gttccactag taccgctcag gtgcagctag ttgaaagcgg cggtgcgctg
      421 gttcagccgg gtggtagcct gcgtctgagc tgtgcggcga gcggctttcc ggtgaaccgt
      481 tatagcatgc gctggtatcg tcaggcaccg ggcaaagaac gtgaatgggt ggcaggcatg
      541 agcagcgcgg gtgatcgtag cagctatgaa gatagcgtga aaggccgttt taccattagc
      601 cgtgatgatg cgcgtaacac cgtgtatctg cagatgaaca gcctgaaacc ggaagatacc
      661 gcggtgtatt attgcaacgt gaacgtgggc tttgaatatt ggggccaggg cacacaggtt
      721 acagttagta gctgacggcc gcacatcacc atcaccatca ctaagcttaa ttagctgagc
      781 ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa
      841 cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aagctagcca tgaaaataaa
      901 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg
      961 tcttgctcta ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     1021 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gattgtatgg gaagcccgat
     1081 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     1141 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     1201 cgtactcctg atgatgcttg gttactcacg actgcgatcc ccggcaaaac agcattccag
     1261 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     1321 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
     1381 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     1441 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcacaaact tttgccattc
     1501 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     1561 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     1621 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     1681 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     1741 gagtttttct aagaattaat tcatgggcaa atattatacg caaggcgaca aggtgctgat
     1801 gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct
     1861 taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag
     1921 ttattggtgc ccttaaacgc ctggggtaat gactctctag cttgaggcat caaataaaac
     1981 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     2041 tcctgagtag gacaaatccg ccgctctaga ttacgtgcag tcgatgataa gctgtcaaac
     2101 atgagaattg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg
     2161 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     2221 gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac
     2281 agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt
     2341 tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg
     2401 tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg
     2461 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
     2521 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
     2581 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
     2641 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
     2701 cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata
     2761 ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca
     2821 gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg
     2881 cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc
     2941 atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt
     3001 tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg
     3061 cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc
     3121 actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc
     3181 tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc
     3241 accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcaccat
     3301 tcgatggtgt cggaatttcg ggcagcgttg ggtcctggcc acgggtgcgc atgatctaga
     3361 gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     3421 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     3481 cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt
     3541 atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     3601 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc
     3661 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3721 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3781 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3841 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3901 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3961 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4021 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4081 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4141 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4201 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4261 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4321 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4381 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4441 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgacattaac
     4501 ctataaaaat aggcgtatca cgaggccctt tcgtcttcac
//
