LOCUS       sequence-403304-        3091 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Level 0 part.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3091)
  AUTHORS   Gilbert C, Brumwell SL, Crits-Christoph A, Kang SC, Martin-Moldes Z,
            Alsharif W, Esmurria A, Nguyen M-A, Lee HH, Ostrov N
  TITLE     A scalable framework for high-throughput identification of 
            functional origins of replication in non-model bacteria
  JOURNAL   bioRxiv 2023.05.19.541510
REFERENCE   2  (bases 1 to 3091)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2023.05.19.541510"
COMMENT     Sequence Label: pGL0_35 [aadA]
FEATURES             Location/Qualifiers
     source          1..3091
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             82..873
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     primer_bind     complement(1135..1151)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1135..1151)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1148..1170)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1323..1911)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1403..1422)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2044..2138)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2159..2818)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2733..2752
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2819..2921)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     3008..3030
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3022..3039
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3023..3039
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tagcgtgcga acagcacgct attgttgtat cattactcgc atccattctc aggctgtctc
       61 gtctcgtctc cgaggtctca aatgagggaa gcggtgatcg ccgaagtatc gactcaacta
      121 tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc cgtacatttg
      181 tacggctctg cagtggatgg cggcctgaag ccacacagtg atattgattt gctggttacg
      241 gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga tcaacgacct tttggaaact
      301 tcggcttccc ctggagagag cgagattctc cgcgctgtag aagtcaccat tgttgtgcac
      361 gacgacatca ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg agaatggcag
      421 cgcaatgaca ttcttgcagg tatcttcgag ccagccacga tcgacattga tctggctatc
      481 ttgctgacaa aagcaagaga acatagcgtt gccttggtag gtccagcggc ggaggaactc
      541 tttgatccgg ttcctgaaca ggatctattt gaggcgctaa atgaaacctt aacgctatgg
      601 aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt
      661 tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg
      721 gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga
      781 caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg
      841 aaaggcgaga tcaccaaggt agtcggcaaa taagctttga gaccacccca ggatacatag
      901 attaccacaa ctccgagccc ttccacctag cgtgctaacc acgcacgcta ttgttgtatc
      961 ccatataatg caatcgtagt ctacctgact gtacttagaa atgtggcttc gcctttgccc
     1021 acgcacctga tcgctcctcg tttgctttta aggaccggac gaaccacaga gcattagaag
     1081 gctaggtgga ggctcagtga tgataagtct gcgatggtgg atgcatgtgt catggtcata
     1141 gctgtttcct gtgtgaaatt gttatccgct cagagggcac aatcctattc cgcgctatcc
     1201 gacaatctcc aagacattag gtggagttca gttcggcgta tggcatatgt cgctggaaag
     1261 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     1321 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     1381 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     1441 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     1501 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     1561 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     1621 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     1681 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     1741 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     1801 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     1861 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     1921 ttgatctttt ctacggggtc tgacgctcta ttcaacaaag ccgccgtccc gtcaagtcag
     1981 cgtaaatggg tagggggctt caaatcgtcc ccccatacga tataagttgt tactagtgct
     2041 tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg
     2101 gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctcg atatcaaatt
     2161 acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat
     2221 ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc
     2281 cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca
     2341 cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct
     2401 caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat
     2461 atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt
     2521 cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac
     2581 cgtctttcat tgccatacga aattccggat gagcattcat caggcgggca agaatgtgaa
     2641 taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat
     2701 ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt
     2761 ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt
     2821 tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta
     2881 tttcattatg gtgaaagttg gaacctctta cgtgccgatc aacgtctcat tgatacctga
     2941 aacaaaaccc atcgtacggc caaggaagtc tccaataact gtgatccacc acaagcgcca
     3001 gggttttccc agtcacgacg ttgtaaaacg acggccagtc atgcataatc cgcacgcatc
     3061 tggaataagg aagtgccatt ccgcctgacc t
//
