LOCUS       sequence-401235-        7618 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 7618)
  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 7618)
  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_3 [RSF1010 ORI]
FEATURES             Location/Qualifiers
     source          1..7618
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     182..201
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(268..370)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     457..479
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     471..488
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     472..488
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(813..1664)
                     /codon_start=1
                     /gene="repC"
                     /product="replication protein C of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepC
                     /translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TADAMVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     CDS             complement(1651..2490)
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepA
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     CDS             complement(3001..3972)
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepB
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     oriT            5211..5298
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      5640..6034
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     primer_bind     complement(6202..6218)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6202..6218)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6215..6237)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(6390..6978)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6470..6489)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(7111..7205)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(7226..7618,1..267))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgaa attccggatg
       61 agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt
      121 ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg
      181 agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt
      241 ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgataa
      301 ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac
      361 gtgccgatca acgtctcatt gatacctgaa acaaaaccca tcgtacggcc aaggaagtct
      421 ccaataactg tgatccacca caagcgccag ggttttccca gtcacgacgt tgtaaaacga
      481 cggccagtca tgcataatcc gcacgcatct ggaataagga agtgccattc cgcctgacct
      541 tagcgtgcga acagcacgct attgttgtat cattactcgc atccattctc aggctgtctc
      601 gtctcgtctc cgaggtctcc ggagatacat ctggaagcaa agccaggaaa gcggcctatg
      661 gagctgtgcg gcagcgctca gtaggcaatt tttcaaaata ttgttaagcc ttttctgagc
      721 atggtatttt tcatggtatt accaattagc aggaaaataa gccattgaat ataaaagata
      781 aaaatgtctt gtttacaata gagtgggggg ggtcagcctg ccgccttggg ccgggtgatg
      841 tcgtacttgc ccgccgcgaa ctcggttacc gtccagccca gcgcgaccag ctccggcaac
      901 gcctcgcgca cccgctggcg gcgcttgcgc atggtcgaac cactggcctc tgacggccag
      961 acatagccgc acaaggtatc tatggaagcc ttgccggttt tgccggggtc gatccagcca
     1021 cacagccgct ggtgcagcag gcgggcggtt tcgctgtcca gcgcccgcac ctcgtccatg
     1081 ctgatgcgca catgctggcc gccacccatg acggcctgcg cgatcaaggg gttcagggcc
     1141 acgtacaggc gcccgtccgc ctcgtcgctg gcgtactccg acagcagccg aaacccctgc
     1201 cgcttgcggc cattctgggc gatgatggat accttccaaa ggcgctcgat gcagtcctgt
     1261 atgtgcttga gcgccccacc actatcgacc tctgccccga tttcctttgc cagcgcccga
     1321 tagctacctt tgaccaccat ggcatcagcg gtgacggcct cccacttggg ttccaggaac
     1381 agccggagct gccgtccgcc ttcggtcttg ggttccgggc caagcactag gccattaggc
     1441 ccagccatgg ccaccagccc ttgcaggatg cgcagatcat cagcgcccag cggctccggg
     1501 ccgctgaact cgatccgctt gccgtcgccg tagtcatacg tcacgtccag cttgctgcgc
     1561 ttgcgctcgc cccgcttgag ggcacggaac aggccggggg ccagacagtg cgccgggtcg
     1621 tgccggacgt ggctgaggct gtgcttgttc ttaggcttca ccacggggca cccccttgct
     1681 cttgcgctgc ctctccagca cggcgggctt gagcaccccg ccgtcatgcc gcctgaacca
     1741 ccgatcagcg aacggtgcgc catagttggc cttgctcaca ccgaagcgga cgaagaaccg
     1801 gcgctggtcg tcgtccacac cccattcctc ggcctcggcg ctggtcatgc tcgacaggta
     1861 ggactgccag cggatgttat cgaccagtac cgagctgccc cggctggcct gctgctggtc
     1921 gcctgcgccc atcatggccg cgcccttgct ggcatggtgc aggaacacga tagagcaccc
     1981 ggtatcggcg gcgatggcct ccatgcgacc gatgacctgg gccatggggc cgctggcgtt
     2041 ttcttcctcg atgtggaacc ggcgcagcgt gtccagcacc atcaggcggc ggccctcggc
     2101 ggcgcgcttg aggccgtcga accactccgg ggccatgatg ttgggcaggc tgccgatcag
     2161 cggctggatc agcaggccgt cagccacggc ttgccgttcc tcggcgctga ggtgcgcccc
     2221 aagggcgtgc aggcggtgat gaatggcggt gggcgggtct tcggcgggca ggtagatcac
     2281 cgggccggtg ggcagttcgc ccacctccag cagatccggc ccgcctgcaa tctgtgcggc
     2341 cagttgcagg gccagcatgg atttaccggc accaccgggc gacaccagcg ccccgaccgt
     2401 accggccacc atgttgggca aaacgtagtc cagcggtggc ggcgctgctg cgaacgcctc
     2461 cagaatattg ataggcttat gggtagccat tgattgcctc ctttgcaggc agttggtggt
     2521 taggcgctgg cggggtcact acccccgccc tgcgccgctc tgagttcttc caggcactcg
     2581 cgcagcgcct cgtattcgtc gtcggtcagc cagaacttgc gctgacgcat ccctttggcc
     2641 ttcatgcgct cggcatatcg cgcttggcgt acagcgtcag ggctggccag caggtcgccg
     2701 gtctgcttgt ccttttggtc tttcatatca gtcaccgaga aacttgccgg ggccgaaagg
     2761 cttgtcttcg cggaacaagg acaaggtgca gccgtcaagg ttaaggctgg ccatatcagc
     2821 gactgaaaag cggccagcct cggccttgtt tgacgtataa ccaaagccac cgggcaacca
     2881 atagcccttg tcacttttga tcaggtagac cgaccctgaa gcgctttttt cgtattccat
     2941 aaaaccccct tctgtgcgtg agtactcata gtataacagg cgtgagtacc aacgcaagca
     3001 ctacatgctg aaatctggcc cgcccctgtc catgcctcgc tggcggggtg ccggtgcccg
     3061 tgccagctcg gcccgcgcaa gctggacgct gggcagaccc atgaccttgc tgacggtgcg
     3121 ctcgatgtaa tccgcttcgt ggccgggctt gcgctctgcc agcgctgggc tggcctcggc
     3181 catggccttg ccgatttcct cggcactgcg gccccggctg gccagcttct gcgcggcgat
     3241 aaagtcgcac ttgctgaggt catcaccgaa gcgcttgacc agcccggcca tctcgctgcg
     3301 gtactcgtcc agcgccgtgc gccggtggcg gctaagctgc cgctcgggca gttcgaggct
     3361 ggccagcctg cgggccttct cctgctgccg ctgggcctgc tcgatctgct ggccagcctg
     3421 ctgcaccagc gccgggccag cggtggcggt cttgcccttg gattcacgca gcagcaccca
     3481 cggctgataa ccggcgcggg tggtgtgctt gtccttgcgg ttggtgaagc ccgccaagcg
     3541 gccatagtgg cggctgtcgg cgctggccgg gtcggcgtcg tactcgctgg ccagcgtccg
     3601 ggcaatctgc ccccgaagtt caccgcctgc ggcgtcggcc accttgaccc atgcctgata
     3661 gttcttcggg ctggtttcca ctaccagggc aggctcccgg ccctcggctt tcatgtcatc
     3721 caggtcaaac tcgctgaggt cgtccaccag caccagacca tgccgctcct gctcggcggg
     3781 cctgatatac acgtcattgc cctgggcatt catccgcttg agccatggcg tgttctggag
     3841 cacttcggcg gctgaccatt cccggttcat catctggccg gtggtggcgt ccctgacgcc
     3901 gatatcgaag cgctcacagc ccatggcctt gagctgtcgg cctatggcct gcaaagtcct
     3961 gtcgttcttc atcgggccac caagcgcagc cagatcgagc cgtcctcggt tgtcagtggc
     4021 gtcaggtcga gcaagagcaa cgatgcgatc agcagcacca ccgtaggcat catggaagcc
     4081 agcatcacgg ttagccatag cttccagtgc cacccccgcg acgcgctccg ggcgctctgc
     4141 gcggcgctgc tcacctcggc ggctacctcc cgcaactctt tggccagctc cacccatgcc
     4201 gcccctgtct ggcgctgggc tttcagccac tccgccgcct gcgcctcgct ggcctgctgg
     4261 gtctggctca tgacctgccg ggcttcgtcg gccagtgtcg ccatgctctg ggccagcggt
     4321 tcgatctgct ccgctaactc gttgatgcct ctggatttct tcactctgtc gattgcgttc
     4381 atggtctatt gcctcccggt attcctgtaa gtcgatgatc tgggcgttgg cggtgtcgat
     4441 gttcagggcc acgtctgccc ggtcggtgcg gatgccccgg ccttccatct ccaccacgtt
     4501 cggccccagg tgaacaccgg gcaggcgctc gatgccctgc gcctcaagtg ttctgtggtc
     4561 aatgcgggcg tcgtggccag cccgctctaa tgcccggttg gcatggtcgg cccatgcctc
     4621 gcgggtctgc tcaagccatg ccttgggctt gagcgcttcg gtcttctgtg ccccgccctt
     4681 ctccggggtc ttgccgttgt accgcttgaa ccactgagcg gcgggccgct cgatgccgtc
     4741 attgatccgc tcggagatca tcaggtggca gtgcgggttc tcgccgccac cggcatggat
     4801 ggccagcgta tacggcaggc gctcggcacc ggtcaggtgc tgggcgaact cggacgccag
     4861 cgccttctgc tggtcgaggg tcagctcgac cggcagggca aattcgacct ccttgaacag
     4921 ccgcccattg gcgcgttcat acaggtcggc agcatcccag tagtcggcgg gccgctcgac
     4981 gaactccggc atgtgcccgg attcggcgtg caagacttca tccatgtcgc gggcatactt
     5041 gccttcgcgc tggatgtagt cggccttggc cctggccgat tggccgcccg acctgctgcc
     5101 ggttttcgcc gtaaggtgat aaatcgccat gctgcctcgc tgttgctttt gcttttcggc
     5161 tccatgcaat ggccctcgga gagcgcaccg cccgaagggt ggccgttagg ccagtttctc
     5221 gaagagaaac cggtaagtgc gccctcccct acaaagtagg gtcgggattg ccgccgctgt
     5281 gcctccatga tagcctacga gacagcacat taacaatggg gtgtcaagat ggttaagggg
     5341 agcaacaagg cggcggatcg gctggccaag ctcgaagaac aacgagcgcg aatcaatgcc
     5401 gaaattcagc gggtgcgggc aagggaacag cagcaagagc gcaagaacga aacaaggcgc
     5461 aaggtgctgg tgggggccat gattttggcc aaggtgaaca gcagcgagtg gccggaggat
     5521 cggctcatgg cggcaatgga tgcgtacctt gaacgcgacc acgaccgcgc cttgttcggt
     5581 ctgccgccac gccagaagga tgagccgggc tgaatgatcg accgagacag gccctgcggg
     5641 gctgcacacg cgcccccacc cttcgggtag ggggaaaggc cgctaaagcg gctaaaagcg
     5701 ctccagcgta tttctgcggg gtttggtgtg gggtttagcg ggctttgccc gcctttcccc
     5761 ctgccgcgca gcggtggggc ggtgtgtagc ctagcgcagc gaatagacca gctatccggc
     5821 ctctggccgg gcatattggg caagggcagc agcgccccac aagggcgctg ataaccgcgc
     5881 ctagtggatt attcttagat aatcatggat ggatttttcc aacaccccgc cagcccccgc
     5941 ccctgctggg tttgcaggtt tgggggcgtg acagttattg caggggttcg tgacagttat
     6001 tgcagggggg cgtgacagtt attgcagggg ttcgtgacag ttagtacggg agtgacgggc
     6061 gctcgagacc accccaggat acatagatta ccacaactcc gagcccttcc acctagcgtg
     6121 ctaaccacgc acgctattgt tgtataggct aggtggaggc tcagtgatga taagtctgcg
     6181 atggtggatg catgtgtcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcag
     6241 agggcacaat cctattccgc gctatccgac aatctccaag acattaggtg gagttcagtt
     6301 cggcgtatgg catatgtcgc tggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6361 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6421 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6481 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6541 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6601 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6661 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6721 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6781 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6841 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6901 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6961 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctctattc
     7021 aacaaagccg ccgtcccgtc aagtcagcgt aaatgggtag ggggcttcaa atcgtccccc
     7081 catacgatat aagttgttac tagtgcttgg attctcacca ataaaaaacg cccggcggca
     7141 accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggat ctatcaacag
     7201 gagtccaagc gagctcgata tcaaattacg ccccgccctg ccactcatcg cagtactgtt
     7261 gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa
     7321 tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg
     7381 gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg
     7441 gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt
     7501 caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt
     7561 attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagg
//
