LOCUS       sequence-401284-        4798 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 4798)
  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 4798)
  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_22 [pSG5 ORI]
FEATURES             Location/Qualifiers
     source          1..4798
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(472..566)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(587..1246)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1161..1180
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1247..1349)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     1436..1458
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1450..1467
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1451..1467
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2115..3557
                     /codon_start=1
                     /gene="rep"
                     /product="replication initiator protein from the 
                     Streptomyces ghanaensis plasmid pSG5"
                     /label=pSG5 Rep
                     /translation="MWKPGEATWGNTCRCNNVHTCPWCMSRILAVRGSNVQLAADGLAD
                     AGYGLHLGTNTLRHFERMAFGTVRKGMRHGLVAVLHDGWKGAYGSSGRRWRTMRDDFGI
                     IGYERAFEDTFGWGSGWHLHWHTLWVTREVLGPDAQAAFRDALAGAWAAGVESAGGYTV
                     SETCDRPGCSCEGKGHGTDVRPLNGADAADGDAGKQARYLYKDGDKTKGGVAKIGLELA
                     GQNFKAGRGDDRMGPLDLGDAAAAELQRLRRPGPFVEKYREREFGVFQVRKHYRSQNLN
                     RLIKELGIQQDVRTEEEITDDTEGLVAIAVIPAYIWYRYIARVAGRRLDLIKVAETYGL
                     PGVRRLVESWGLVWGKDVLDPPAPEAPAAPGDLDADQMRFEVMSEEEAAFREARRKANE
                     ARTEELAASLDRVRQPKKEAIRPTISLRKRLKPKPVTVDVKTPPPGAASPVCRRCKGKL
                     APVLQPWGQHPGDCLRVDTAVA"
     primer_bind     complement(4361..4377)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4361..4377)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4374..4396)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(join(4549..4798,1..339))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4629..4648)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
       61 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
      121 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
      181 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
      241 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
      301 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
      361 acggggtctg acgctctatt caacaaagcc gccgtcccgt caagtcagcg taaatgggta
      421 gggggcttca aatcgtcccc ccatacgata taagttgtta ctagtgcttg gattctcacc
      481 aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga gttctgaggt
      541 cattactgga tctatcaaca ggagtccaag cgagctcgat atcaaattac gccccgccct
      601 gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac
      661 aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat
      721 atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa
      781 aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt
      841 tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa
      901 actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat
      961 ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg
     1021 ccatacgaaa ttccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat
     1081 aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg
     1141 tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc
     1201 attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag
     1261 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt
     1321 gaaagttgga acctcttacg tgccgatcaa cgtctcattg atacctgaaa caaaacccat
     1381 cgtacggcca aggaagtctc caataactgt gatccaccac aagcgccagg gttttcccag
     1441 tcacgacgtt gtaaaacgac ggccagtcat gcataatccg cacgcatctg gaataaggaa
     1501 gtgccattcc gcctgacctt agcgtgcgaa cagcacgcta ttgttgtatc attactcgca
     1561 tccattctca ggctgtctcg tctcgtctcc gaggtctccg gaggatcaac aggacccaga
     1621 aatggcacga gcccgggatt ggcctcccgg gctcgccgcc cccgaaccgg cgcagcatgc
     1681 cgagtgagga cgtttgctct gtgacgagcg tacgagagaa cagcgcgtcg acaccaccgc
     1741 gccccgccgg accgaagtgc cactgcggca ccccgatcgt ccacatcccc gggaagcgcc
     1801 ccaaaatcta ctgctcggac gcctgccgaa agcgcgcaaa gcgtctcatt gcggaccagt
     1861 cacgcagcgt agcaagcacg actacagcgg gtaatccgga cgcgggcttt agtagtaagg
     1921 gaaaatctcc gtccactgca ggtcagagcg ctgtaactgg aacgggttct accccaaacg
     1981 agggtgccgc gacagccggg cgggaggacc cgcgcgacga gcggttcgcc cggcgggacc
     2041 ggcaccagac ggtgtcgctg aacgaggcgt tcaagggctg cgggacccgg ctcactgccg
     2101 ggacggccga gctgatgtgg aagccgggcg aggcgacgtg gggcaacacg tgccgctgca
     2161 acaacgtcca cacctgcccg tggtgcatga gccgcatcct ggccgtacgg ggctcgaacg
     2221 tgcagctcgc cgccgacggc ctcgcggacg ccgggtacgg gctgcacctt ggcacgaaca
     2281 cgttgcgcca cttcgagcgc atggcgttcg gcaccgtccg caagggaatg cggcacggcc
     2341 tggtcgcggt cttgcacgac ggctggaagg gcgcgtacgg ctcgtccggc cgccgctggc
     2401 gcacgatgcg cgacgacttc gggatcatcg gctacgagcg ggccttcgag gacaccttcg
     2461 gctggggctc cggctggcac ctgcactggc acacgctctg ggtcacccgc gaggtgctcg
     2521 gcccggacgc ccaggccgcg ttccgcgacg cactcgccgg agcctgggcc gccggggtcg
     2581 aatccgcggg cggctacacc gtcagcgaga cgtgcgaccg gcccggttgc tcctgtgagg
     2641 gcaagggcca cggcaccgac gtgcgcccac tcaacggcgc ggacgccgct gacggcgatg
     2701 cgggcaagca ggcccgctac ctctacaagg acggcgacaa gaccaagggc ggcgtcgcca
     2761 agatcggcct ggagctggcc ggacagaact tcaaggccgg tcgcggcgac gaccgcatgg
     2821 gaccgctcga cctgggcgac gcggcggccg ccgagctgca gcggcttcgt cggcccggcc
     2881 cgttcgtcga gaagtaccgc gagcgcgagt tcggcgtctt ccaggtccgc aagcactacc
     2941 ggtcgcagaa cctgaaccgg ctgatcaagg aactcggcat ccagcaggac gtccgcaccg
     3001 aggaggagat caccgacgac accgagggcc tggtcgccat cgcggtcatc ccggcctaca
     3061 tctggtaccg atacatcgcc cgcgtcgccg gacgtcgcct cgacctcatc aaggtcgccg
     3121 agacgtacgg cctaccgggc gtccggcggc tcgtcgagtc ctggggcctg gtgtggggca
     3181 aggacgtcct ggacccgccc gccccggaag cgccggccgc gccgggcgac cttgacgctg
     3241 accagatgag gttcgaggtc atgtccgagg aggaagctgc tttccgcgag gcgcgccgga
     3301 aggccaacga ggcccgcacg gaggagctgg ccgcttcgct cgaccgagtg cgtcagccga
     3361 agaaggaggc gatccgcccg acgatcagcc ttcggaagcg cctgaagccg aagccggtca
     3421 ccgtcgacgt caagactccc ccgcccggcg ccgcgtcgcc cgtgtgccgc cgctgcaagg
     3481 gcaagctcgc cccggtactc cagccctggg gccagcaccc cggagactgc ctccgcgtcg
     3541 acacagcggt cgcgtagggc cgctgaggga ccgccatcag accgccaccc cggacatccg
     3601 ggtggcggtc tgatggcggt cccggcgcga aagccgaggg tctgcctgcc gtgaggtggc
     3661 cggcggacct gggttactgc tggaatcccg cgcccagtcc gagcgtccgc cggcgctttt
     3721 cacctggctc cgggtgctcg tcgatgagca ccaggccgtc gacgacgacg cgtccgcctg
     3781 cctgcttttg gccggcgggc cgggccgcgg ttgtccgccg ggtcggaccg gtcccttggt
     3841 cgaccgatgc ctcggccacg acgatgctgg gcccgaagcg gtacgcgacc cgtgccgggc
     3901 cgcagcggat cgcggcagca ggagctcccc gcccgccacg gctggctgga tggtcgcctc
     3961 acgcgcccgc aggggtgacg gcgtgggtgc ggcggtcagc cccgtgatcg ggcgtcagac
     4021 ggccgctgag ggaccgcgac cacccctccc ttcccgcctg ccgtctttca ccgcccggcc
     4081 ggtccgcaga cgccgtacgg ccaggccacg gaggattacg ggcagatggc gcccacccgg
     4141 cacccgggct ggggcaccct ggacagcacg aaggtccgcc ccactcaccg gggcgggcct
     4201 tcgcgttgag ctagagttcg ctcgagacca ccccaggata catagattac cacaactccg
     4261 agcccttcca cctagcgtgc taaccacgca cgctattgtt gtataggcta ggtggaggct
     4321 cagtgatgat aagtctgcga tggtggatgc atgtgtcatg gtcatagctg tttcctgtgt
     4381 gaaattgtta tccgctcaga gggcacaatc ctattccgcg ctatccgaca atctccaaga
     4441 cattaggtgg agttcagttc ggcgtatggc atatgtcgct ggaaagaaca tgtgagcaaa
     4501 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4561 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4621 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4681 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4741 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggc
//
