LOCUS       sequence-393206-        5217 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 part.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5217)
  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 5217)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 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_12 [pAM-beta-1 ORI]
FEATURES             Location/Qualifiers
     source          1..5217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(3261..3277)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3261..3277)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3274..3296)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(3449..4037)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3529..3548)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4170..4264)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4285..4944)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4859..4878
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4945..5047)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     5134..5156
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5148..5165
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5149..5165
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tagcgtgcga acagcacgct attgttgtat cattactcgc atccattctc aggctgtctc
       61 gtctcgtctc cgaggtctca ggagcccaag aattagaaat gagtagatca aattattcac
      121 gaatagaatc aggaaaatca gatccaacca taaaaacact agaacaaatt gcaaagttaa
      181 ctaactcaac gctagtagtg gatttaatcc caaatgagcc aacagaacca gaaccagaaa
      241 cagaatcaga acaagtaaca ttggatttag aaatggaaga agaaaaaagc aatgacttcg
      301 tgtgaataat gcacgaaatc gttgcttatt ttttttaaaa gcggtatact agatataacg
      361 aaacaacgaa ctgaatagaa acgaaaaaag agccatgaca catttataaa atgtttgacg
      421 acattttata aatgcatagc ccgataagat tgccaaacca acgcttatca gttagtcaga
      481 tgaactcttc cctcgtaaga agttatttaa ttaactttgt ttgaagacgg tatataaccg
      541 tactatcatt atatagggaa atcagagagt tttcaagtat ctaagctact gaatttaaga
      601 attgttaagc aatcaatcgg aaatcgtttg attgcttttt ttgtattcat ttatagaagg
      661 tggagtttgt atgaatcatg atgaatgtaa aacttatata aaaaatagtt tattggagat
      721 aagaaaatta gcaaatatct atacactaga aacgtttaag aaagagttag aaaagagaaa
      781 tatctactta gaaacaaaat cagataagta tttttcttcg gagggggaag attatatata
      841 taagttaata gaaaataaca aaataattta ttcgattagt ggaaaaaaat tgacttataa
      901 aggaaaaaaa tctttttcaa aacatgcaat attgaaacag ttgaatgaaa aagcaaacca
      961 agttaattaa acaacctatt ttataggatt tataggaaag gagaacagct gaatgaatat
     1021 cccttttgtt gtagaaactg tgcttcatga cggcttgtta aagtacaaat ttaaaaatag
     1081 taaaattcgc tcaatcacta ccaagccagg taaaagcaaa ggggctattt ttgcgtatcg
     1141 ctcaaaatca agcatgattg gcggtcgtgg tgttgttctg acttccgagg aagcgattca
     1201 agaaaatcaa gatacattta cacattggac acccaacgtt tatcgttatg gaacgtatgc
     1261 agacgaaaac cgttcataca cgaaaggaca ttctgaaaac aatttaagac aaatcaatac
     1321 cttctttatt gattttgata ttcacacggc aaaagaaact atttcagcaa gcgatatttt
     1381 aacaaccgct attgatttag gttttatgcc tactatgatt atcaaatctg ataaaggtta
     1441 tcaagcatat tttgttttag aaacgccagt ctatgtgact tcaaaatcag aatttaaatc
     1501 tgtcaaagca gccaaaataa tttcgcaaaa tatccgagaa tattttggaa agtctttgcc
     1561 agttgatcta acgtgtaatc attttggtat tgctcgcata ccaagaacgg acaatgtaga
     1621 attttttgat cctaattacc gttattcttt caaagaatgg caagattggt ctttcaaaca
     1681 aacagataat aagggcttta ctcgttcaag tctaacggtt ttaagcggta cagaaggcaa
     1741 aaaacaagta gatgaaccct ggtttaatct cttattgcac gaaacgaaat tttcaggaga
     1801 aaagggttta atagggcgta ataacgtcat gtttaccctc tctttagcct actttagttc
     1861 aggctattca atcgaaacgt gcgaatataa tatgtttgag tttaataatc gattagatca
     1921 acccttagaa gaaaaagaag taatcaaaat tgttagaagt gcctattcag aaaactatca
     1981 aggggctaat agggaataca ttaccattct ttgcaaagct tgggtatcaa gtgatttaac
     2041 cagtaaagat ttatttgtcc gtcaagggtg gtttaaattc aagaaaaaaa gaagcgaacg
     2101 tcaacgtgtt catttgtcag aatggaaaga agatttaatg gcttatatta gcgaaaaaag
     2161 cgatgtatac aagccttatt tagtgacgac caaaaaagag attagagaag tgctaggcat
     2221 tcctgaacgg acattagata aattgctgaa ggtactgaag gcgaatcagg aaattttctt
     2281 taagattaaa ccaggaagaa atggtggcat tcaacttgct agtgttaaat cattgttgct
     2341 atcgatcatt aaagtaaaaa aagaagaaaa agaaagctat ataaaggcgc tgacaaattc
     2401 ttttgactta gagcatacat tcattcaaga gactttaaac aagctagcag aacgccctaa
     2461 aacggacaca caactcgatt tgtttagcta tgatacaggc tgaaaataaa acccgcacta
     2521 tgccattaca tttatatcta tgatacgtgt ttgttttttc tttgctgttt agcgaatgat
     2581 tagcagaaat atacagagta agattttaat taattattag ggggagaagg agagagtagc
     2641 ccgaaaactt ttagttggct tggactgaac gaagtgaggg aaaggctact aaaacgtcga
     2701 ggggcagtga gagcgaagcg aacacttgat tttttaattt tctatctttt ataggtcatt
     2761 agagtatact tatttgtcct ataaactatt tagcagcata atagatttat tgaataggtc
     2821 atttaagttg agcatattag aggaggaaaa tcttggagaa atatttgaag aacccgatta
     2881 catggattgg attagttctt gtggttacgt ggtttttaac taaaagtagt gaatttttga
     2941 tttttggtgt gtgtgtcttg ttgttagtat ttgctagtca aagtgattaa atagaattcc
     3001 gcttgagacc accccaggat acatagatta ccacaactcc gagcccttcc acctagcgtg
     3061 ctaaccacgc acgctattgt tgtatcccat ataatgcaat cgtagtctac ctgactgtac
     3121 ttagaaatgt ggcttcgcct ttgcccacgc acctgatcgc tcctcgtttg cttttaagga
     3181 ccggacgaac cacagagcat tagaaggcta ggtggaggct cagtgatgat aagtctgcga
     3241 tggtggatgc atgtgtcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaga
     3301 gggcacaatc ctattccgcg ctatccgaca atctccaaga cattaggtgg agttcagttc
     3361 ggcgtatggc atatgtcgct ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3421 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3481 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3541 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3601 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3661 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3721 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3781 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3841 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3901 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3961 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4021 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctctattca
     4081 acaaagccgc cgtcccgtca agtcagcgta aatgggtagg gggcttcaaa tcgtcccccc
     4141 atacgatata agttgttact agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa
     4201 ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg
     4261 agtccaagcg agctcgatat caaattacgc cccgccctgc cactcatcgc agtactgttg
     4321 taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat
     4381 cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg
     4441 ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg
     4501 attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc
     4561 accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta
     4621 ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg
     4681 aacactatcc catatcacca gctcaccgtc tttcattgcc atacgaaatt ccggatgagc
     4741 attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt
     4801 tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc
     4861 aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt
     4921 atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc tcgataactc
     4981 aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg
     5041 ccgatcaacg tctcattgat acctgaaaca aaacccatcg tacggccaag gaagtctcca
     5101 ataactgtga tccaccacaa gcgccagggt tttcccagtc acgacgttgt aaaacgacgg
     5161 ccagtcatgc ataatccgca cgcatctgga ataaggaagt gccattccgc ctgacct
//
