LOCUS       sequence-393208-        4448 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 4448)
  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 4448)
  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_13 [pIP501 ORI]
FEATURES             Location/Qualifiers
     source          1..4448
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2492..2508)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2492..2508)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2505..2527)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(2680..3268)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2760..2779)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3401..3495)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3516..4175)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4090..4109
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4176..4278)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     4365..4387
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4379..4396
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4380..4396
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tagcgtgcga acagcacgct attgttgtat cattactcgc atccattctc aggctgtctc
       61 gtctcgtctc cgaggtctca ggagaactaa ctcaacgcta gtagtggatt taatcccaaa
      121 tgagccaaca gaaccagaac cagaaacaga acaagtaaca ttggagttag aaatggaaga
      181 agaaaaaagc aatgatttcg tgtgaataat gcacgaaatc attgcttatt tttttaaaaa
      241 gcgatatact agatataacg aaacaacgaa ctgaataaag aatacaaaaa aagagccacg
      301 accagttaaa gcctgagaaa ttttaactgc gagccttaat tgattaccac caatcaatta
      361 aagaagtcca gacccaaaat ttggtaaagt atttaattac tttattaatc agatacttaa
      421 atatctgtaa acccattata tcgggttttt gaggggattt caagtcttta agaagatacc
      481 aggcaatcaa ttaagaaaaa cttagttgat tgcctttttt gttgtgattc aactttgatc
      541 gtagcttcta actaattaat tttcgtaaga aaggagaaca gctgaatgaa tatccctttt
      601 gttgtagaaa ctgtgcttca tgacggcttg ttaaagtaca aatttaaaaa tagtaaaatt
      661 cgctcaatca ctaccaagcc aggtaaaagt aaaggggcta tttttgcgta tcgctcaaaa
      721 aaaagcatga ttggcggacg tggcgttgtt ctgacttccg aagaagcgat tcacgaaaat
      781 caagatacat ttacgcattg gacaccaaac gtttatcgtt atggtacgta tgcagacgaa
      841 aaccgttcat acactaaagg acattctgaa aacaatttaa gacaaatcaa taccttcttt
      901 attgattttg atattcacac ggaaaaagaa actatttcag caagcgatat tttaacaaca
      961 gctattgatt taggttttat gcctacgtta attatcaaat ctgataaagg ttatcaagca
     1021 tattttgttt tagaaacgcc agtctatgtg acttcaaaat cagaatttaa atctgtcaaa
     1081 gcagccaaaa taatctcgca aaatatccga gaatattttg gaaagtcttt gccagttgat
     1141 ctaacgtgca atcattttgg gattgctcgt ataccaagaa cggacaatgt agaatttttt
     1201 gatcccaatt accgttattc tttcaaagaa tggcaagatt ggtctttcaa acaaacagat
     1261 aataagggct ttactcgttc aagtctaacg gttttaagcg gtacagaagg caaaaaacaa
     1321 gtagatgaac cctggtttaa tctcttattg cacgaaacga aattttcagg agaaaagggt
     1381 ttagtagggc gcaatagcgt tatgtttacc ctctctttag cctactttag ttcaggctat
     1441 tcaatcgaaa cgtgcgaata taatatgttt gagtttaata atcgattaga tcaaccctta
     1501 gaagaaaaag aagtaatcaa aattgttaga agtgcctatt cagaaaacta tcaaggggct
     1561 aatagggaat acattaccat tctttgcaaa gcttgggtat caagtgattt aaccagtaaa
     1621 gatttatttg tccgtcaagg gtggtttaaa ttcaagaaaa aaagaagcga acgtcaacgt
     1681 gttcatttgt cagaatggaa agaagattta atggcttata ttagcgaaaa aagcgatgta
     1741 tacaagcctt atttagcgac gaccaaaaaa gagattagag aagtgctagg cattcctgaa
     1801 cggacattag ataaattgct gaaggtactg aaggcgaatc aggaaatttt ctttaagatt
     1861 aaaccaggaa gaaatggtgg cattcaactt gctagtgtta aatcattgtt gctatcgatc
     1921 attaaattaa aaaaagaaga acgagaaagc tatataaagg cgctgacagc ttcgtttaat
     1981 ttagaacgta catttattca agaaactcta aacaaattgg cagaacgccc caaaacggac
     2041 ccacaactcg atttgtttag ctacgataca ggctgaaaat aaaacccgca ctatgccatt
     2101 acatttatat ctatgatacg tgtttgtttt tctttgctgt ttagtgaatg attagcagaa
     2161 atatacagag taagatttta attaattatt agggggagaa ggagagagta gcccgaaaac
     2221 ttttagttgg cgcttgagac caccccagga tacatagatt accacaactc cgagcccttc
     2281 cacctagcgt gctaaccacg cacgctattg ttgtatccca tataatgcaa tcgtagtcta
     2341 cctgactgta cttagaaatg tggcttcgcc tttgcccacg cacctgatcg ctcctcgttt
     2401 gcttttaagg accggacgaa ccacagagca ttagaaggct aggtggaggc tcagtgatga
     2461 taagtctgcg atggtggatg catgtgtcat ggtcatagct gtttcctgtg tgaaattgtt
     2521 atccgctcag agggcacaat cctattccgc gctatccgac aatctccaag acattaggtg
     2581 gagttcagtt cggcgtatgg catatgtcgc tggaaagaac atgtgagcaa aaggccagca
     2641 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2701 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2761 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2821 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2881 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2941 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3001 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3061 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3121 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3181 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3241 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3301 cgctctattc aacaaagccg ccgtcccgtc aagtcagcgt aaatgggtag ggggcttcaa
     3361 atcgtccccc catacgatat aagttgttac tagtgcttgg attctcacca ataaaaaacg
     3421 cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggat
     3481 ctatcaacag gagtccaagc gagctcgata tcaaattacg ccccgccctg ccactcatcg
     3541 cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga
     3601 tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg
     3661 gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa
     3721 ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag
     3781 gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa
     3841 tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg
     3901 taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc catacgaaat
     3961 tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc
     4021 ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag
     4081 gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata
     4141 tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat
     4201 ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa
     4261 cctcttacgt gccgatcaac gtctcattga tacctgaaac aaaacccatc gtacggccaa
     4321 ggaagtctcc aataactgtg atccaccaca agcgccaggg ttttcccagt cacgacgttg
     4381 taaaacgacg gccagtcatg cataatccgc acgcatctgg aataaggaag tgccattccg
     4441 cctgacct
//
