LOCUS       sequence-385360-        4405 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 4405)
  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 4405)
  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_1 [RK2 ORI]
FEATURES             Location/Qualifiers
     source          1..4405
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     237..256
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(323..425)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     512..534
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     526..543
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     527..543
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(730..1878)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      2271..2802
                     /label=oriV
                     /note="incP origin of replication"
     primer_bind     complement(3044..3060)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3044..3060)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3057..3079)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(3232..3820)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3312..3331)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3953..4047)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(4068..4405,1..322))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa
       61 cactatccca tatcaccagc tcaccgtctt tcattgccat acgaaattcc ggatgagcat
      121 tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta
      181 cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa
      241 ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat
      301 atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa
      361 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
      421 gatcaacgtc tcattgatac ctgaaacaaa acccatcgta cggccaagga agtctccaat
      481 aactgtgatc caccacaagc gccagggttt tcccagtcac gacgttgtaa aacgacggcc
      541 agtcatgcat aatccgcacg catctggaat aaggaagtgc cattccgcct gaccttagcg
      601 tgcgaacagc acgctattgt tgtatcatta ctcgcatcca ttctcaggct gtctcgtctc
      661 gtctccgagg tctccggagg cgatgcaggt ggctgctgaa cccccagccg gaactgaccc
      721 cacaaggccc tagcgtttgc aatgcaccag gtcatcattg acccaggcgt gttccaccag
      781 gccgctgcct cgcaactctt cgcaggcttc gccgacctgc tcgcgccact tcttcacgcg
      841 ggtggaatcc gatccgcaca tgaggcggaa ggtttccagc ttgagcgggt acggctcccg
      901 gtgcgagctg aaatagtcga acatccgtcg ggccgtcggc gacagcttgc ggtacttctc
      961 ccatatgaat ttcgtgtagt ggtcgccagc aaacagcacg acgatttcct cgtcgatcag
     1021 gacctggcaa cgggacgttt tcttgccacg gtccaggacg cggaagcggt gcagcagcga
     1081 caccgattcc aggtgcccaa cgcggtcgga cgtgaagccc atcgccgtcg cctgtaggcg
     1141 cgacaggcat tcctcggcct tcgtgtaata ccggccattg atcgaccagc ccaggtcctg
     1201 gcaaagctcg tagaacgtga aggtgatcgg ctcgccgata ggggtgcgct tcgcgtactc
     1261 caacacctgc tgccacacca gttcgtcatc gtcggcccgc agctcgacgc cggtgtaggt
     1321 gatcttcacg tccttgttga cgtggaaaat gaccttgttt tgcagcgcct cgcgcgggat
     1381 tttcttgttg cgcgtggtga acagggcaga gcgggccgtg tcgtttggca tcgctcgcat
     1441 cgtgtccggc cacggcgcaa tatcgaacaa ggaaagctgc atttccttga tctgctgctt
     1501 cgtgtgtttc agcaacgcgg cctgcttggc ttcgctgacc tgttttgcca ggtcctcgcc
     1561 ggcggttttt cgcttcttgg tcgtcatagt tcctcgcgtg tcgatggtca tcgacttcgc
     1621 caaacctgcc gcctcctgtt cgagacgacg cgaacgctcc acggcggccg atggcgcggg
     1681 cagggcaggg ggagccagtt gcacgctgtc gcgctcgatc ttggccgtag cttgctggac
     1741 tatcgagccg acggactgga aggtttcgcg gggcgcacgc atgacggtgc ggcttgcgat
     1801 ggtttcggca tcctcggcgg aaaaccccgc gtcgatcagt tcttgcctgt atgccttccg
     1861 gtcaaacgtc cgattcattc accctccttg cgggattgcc ccggaattaa ttccccggat
     1921 cgatccgtcg atcttgatcc cctgcgccat cagatccttg gcggcaagaa agccatccag
     1981 tttactttgc agggcttccc aaccttacca gagggcgccc cagctggcaa ttccggttcg
     2041 cttgctgtcc ataaaaccgc ccagtctagc tatcgccatg taagcccact gcaagctacc
     2101 tgctttctct ttgcgcttgc gttttccctt gtccagatag cccagtagct gacattcatc
     2161 cggggtcagc accgtttctg cggactggct ttctacgtgg ctgccatttt tggggtgagg
     2221 ccgttcgcgg ccgaggggcg cagcccctgg ggggatggga ggcccgcgtt agcgggccgg
     2281 gagggttcga gaaggggggg cacccccctt cggcgtgcgc ggtcacgcgc acagggcgca
     2341 gccctggtta aaaacaaggt ttataaatat tggtttaaaa gcaggttaaa agacaggtta
     2401 gcggtggccg aaaaacgggc ggaaaccctt gcaaatgctg gattttctgc ctgtggacag
     2461 cccctcaaat gtcaataggt gcgcccctca tctgtcagca ctctgcccct caagtgtcaa
     2521 ggatcgcgcc cctcatctgt cagtagtcgc gcccctcaag tgtcaatacc gcagggcact
     2581 tatccccagg cttgtccaca tcatctgtgg gaaactcgcg taaaatcagg cgttttcgcc
     2641 gatttgcgag gctggccagc tccacgtcgc cggccgaaat cgagcctgcc cctcatctgt
     2701 caacgccgcg ccgggtgagt cggcccctca agtgtcaacg tccgcccctc atctgtcagt
     2761 gagggccaag ttttccgcga ggtatccaca acgccggcgg ccctacatgg ctctgctgta
     2821 gtgagtgggt tgcgctccgg cagcggtcct gatcccccgc agaaaaaaag gatctcaaga
     2881 agatcctttg atcttttcta ccgctcgaga ccaccccagg atacatagat taccacaact
     2941 ccgagccctt ccacctagcg tgctaaccac gcacgctatt gttgtatagg ctaggtggag
     3001 gctcagtgat gataagtctg cgatggtgga tgcatgtgtc atggtcatag ctgtttcctg
     3061 tgtgaaattg ttatccgctc agagggcaca atcctattcc gcgctatccg acaatctcca
     3121 agacattagg tggagttcag ttcggcgtat ggcatatgtc gctggaaaga acatgtgagc
     3181 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3241 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3301 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3361 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3421 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3481 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3541 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3601 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3661 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3721 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3781 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3841 tacggggtct gacgctctat tcaacaaagc cgccgtcccg tcaagtcagc gtaaatgggt
     3901 agggggcttc aaatcgtccc cccatacgat ataagttgtt actagtgctt ggattctcac
     3961 caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg
     4021 tcattactgg atctatcaac aggagtccaa gcgagctcga tatcaaatta cgccccgccc
     4081 tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca
     4141 caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa
     4201 tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca
     4261 aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct
     4321 ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga
     4381 aactgccgga aatcgtcgtg gtatt
//
