LOCUS       sequence-401232-        4031 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 4031)
  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 4031)
  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_110 [pNG2]
FEATURES             Location/Qualifiers
     source          1..4031
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(57..73)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(57..73)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(70..92)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(245..833)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(325..344)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(966..1060)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1081..1740)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1655..1674
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1741..1843)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     1930..1952
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1944..1961
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1945..1961
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aggctaggtg gaggctcagt gatgataagt ctgcgatggt ggatgcatgt gtcatggtca
       61 tagctgtttc ctgtgtgaaa ttgttatccg ctcagagggc acaatcctat tccgcgctat
      121 ccgacaatct ccaagacatt aggtggagtt cagttcggcg tatggcatat gtcgctggaa
      181 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
      241 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
      301 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
      361 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
      421 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
      481 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
      541 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
      601 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
      661 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
      721 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
      781 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
      841 ctttgatctt ttctacgggg tctgacgctc tattcaacaa agccgccgtc ccgtcaagtc
      901 agcgtaaatg ggtagggggc ttcaaatcgt ccccccatac gatataagtt gttactagtg
      961 cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga
     1021 tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct cgatatcaaa
     1081 ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac
     1141 atggaagcca tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc
     1201 gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc
     1261 cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt
     1321 ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga
     1381 atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt
     1441 ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc
     1501 accgtctttc attgccatac gaaattccgg atgagcattc atcaggcggg caagaatgtg
     1561 aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat
     1621 atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg
     1681 ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat
     1741 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct
     1801 tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaacgtctc attgatacct
     1861 gaaacaaaac ccatcgtacg gccaaggaag tctccaataa ctgtgatcca ccacaagcgc
     1921 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tcatgcataa tccgcacgca
     1981 tctggaataa ggaagtgcca ttccgcctga ccttagcgtg cgaacagcac gctattgttg
     2041 tatcattact cgcatccatt ctcaggctgt ctcgtctcgt ctccgaggtc tcaggagatg
     2101 gtaaatctgc gcagacagcc ctgtgcagct gaaacgcggt tacgtatagc ttgccatatg
     2161 tctagccata cgtaaccgca ggtaaaaggc atatttttcg cgtgtcatgg ctagtaaata
     2221 acaccggtgt catttagagt cagggaaaga caatgaaaaa cgaagaaagc caccgggcgg
     2281 caacccgatg actttcgctt atcacccagc acacacctgg gagaaatcac ggtcatgagt
     2341 ttacagactc atgcgcagaa tgcgcacact aaaacaccta cccgcgtcga gcgcgaccgt
     2401 ggtggactgg acaacacccc agcatctgcc agtgaccgcg accttttacg cgatcatcta
     2461 ggccgcgatg tactccacgg ttcagtcaca cgagacttta aaaaggccta tcgacgcaac
     2521 gctgacggca cgaactcgcc gcgtatgtat cgcttcgaga ctgatgcttt aggacggtgc
     2581 gagtacgcca tgctcaccac caagcagtac gccgccgtcc tggtcgtaga cgttgaccaa
     2641 gtaggtaccg caggcggtga ccccgcagac ttaaacccgt acgtccgcga cgtggtgcgc
     2701 tcactgatta ctcatagcgt cgggccagcc tgggtgggta ttaacccaac taacggcaaa
     2761 gcccagttca tatggcttat tgaccctgtc tacgctgacc gtaacggtaa atctgcgcag
     2821 atgaagcttc ttgcagcaac cacgcgtgtg ctgggtgagc ttttagacca tgacccgcac
     2881 ttttcccacc gctttagccg caacccgttc tacacaggca aagcccctac cgcttatcgt
     2941 tggtataggc agcacaaccg ggtgatgcgc cttggagact tgataaagca ggtaagggat
     3001 atggcaggac acgaccagtt caaccccacc ccacgccagc aattcagctc tggccgcgaa
     3061 cttatcaacg cggtcaagac ccgccgtgaa gaagcccaag cattcaaagc actcgcccag
     3121 gacgtagacg cggaaatcgc cggtggcctc gaccagtatg acccggaact tatcgacggt
     3181 gtgcgtgtgc tctggattgt ccaaggaacc gcagcacgcg acgaaacagc ctttagacat
     3241 gcgcttaaga ctggccaccg cttgcgccag caaggccaac gcctgacaga cgcagcaatc
     3301 atcgacgcct atgagcacgc ctacaacgtc gcacacaccc acggcggtgc aggccgcgac
     3361 aacgagatgc cacccatgcg cgaccgccaa accatggcaa ggcgcgtgcg cgggtatgtc
     3421 gcccaatcca agagcgaaac ctacagcggc tctaacgcac caggtaaagc caccagcagc
     3481 gagcggaaag ccttggccac gatgggacgc agaggcggac aaaaagccgc acaacgctgg
     3541 aaaacagacc ccgagggcaa atatgcgcaa gcacaaaggt cgaagcttga aaagacgcac
     3601 cgtaagaaaa aggctcaagg acgatctacg aagtcccgta ttagccaaat ggtgaacgat
     3661 cagtatttcc agacagggac agttcccacg tgggctgaaa taggggcaga ggtaggagtc
     3721 tctcgcgcca cggttgctag gcatgtcgcg gagctaaaga aaagcggtga ctatccggac
     3781 gtttaaggcg tctcataccg taagcaatat acggttcccc tgccgttagg cagttagata
     3841 aaacctcact tgaagaaaac cttgaggggc agggcagctt atatgcttca aagcatgact
     3901 tcctctgttc tcctagacct cgcaaccctc cgccataacc tcacccgctt gagaccaccc
     3961 caggatacat agattaccac aactccgagc ccttccacct agcgtgctaa ccacgcacgc
     4021 tattgttgta t
//
