LOCUS       sequence-397974-        3219 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 3219)
  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 3219)
  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_143 [EilR-PjexH1]
FEATURES             Location/Qualifiers
     source          1..3219
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             799..807
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(1263..1279)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1263..1279)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1276..1298)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1451..2039)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1531..1550)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2172..2266)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2287..2946)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2861..2880
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2947..3049)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     3136..3158
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3150..3167
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3151..3167
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tagcgtgcga acagcacgct attgttgtat cattactcgc atccattctc aggctgtctc
       61 gtctcgtctc cgaggtctca ggagggcccg acgcgcgccc agtcgacggg gaatttgccg
      121 agatggggga ccggcgcgag cgcgtagaag gtcgaatggc ccggcggcgc catttttggg
      181 tcggtcgcgg tcgggtgatg gagatagagc gagttacgaa aataactcaa gctgaataac
      241 gtgctgcagg tggcgggtga atgcggcatc gtctacctcc ggcattccca gcacgtaaat
      301 cccgtccagt ccacagacca gcgaaatcag ccgccaggcg atattttcgg cgctatcgcg
      361 cagggtaaat tcgccagcgg catggcccgc gcgaatgatc ctgaccgctt cgtcatgcca
      421 caggttcatg gtcagcaggt acgcgctttt gatttccgga tcgctgtcgg ccagcagctg
      481 cgcctggcgc cacaggcgga tatagggctc cagccgtccg tcctcgctgc cgagggcgga
      541 aaacagctgt tcacgccagc cggcggtgcg cgaaaggcgc tgcagatcca tcatctcgcg
      601 gatcacgcgg ataaacgcct gggattttag ctcgccggag gaggtgaaat ggtgatggac
      661 ctggcctgcg gcaacacccg ctgcggtggc gatgttgcgg accgtcatac cggtaaaccc
      721 ttgatccagc gcgacgcgca tggcggcctg catgatggtt tcccgacgtt cttcgcgatt
      781 cagatagccc atattgtcac ctccttatcc acacattata cgagccggat gattaatggg
      841 gaaccttacc agagggcgcc ccagctggca attcctctta agtaagtaag agtatacgta
      901 tatcggctaa taacgtatta aggcgcttcg gcgccttttt ttatgggggt attttcatcc
      961 caatccacac gtccaacgca cagcaaacac cacgtcgacc ctatcagctg cgtgctttct
     1021 atgagtcgtt gctgcataac ataattttta aaaaattcat ttgctaaagt tggacacgtg
     1081 tcctataata tacttcaaag ttggacacgt gtccaacttt tacttgagac caccccagga
     1141 tacatagatt accacaactc cgagcccttc cacctagcgt gctaaccacg cacgctattg
     1201 ttgtataggc taggtggagg ctcagtgatg ataagtctgc gatggtggat gcatgtgtca
     1261 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca gagggcacaa tcctattccg
     1321 cgctatccga caatctccaa gacattaggt ggagttcagt tcggcgtatg gcatatgtcg
     1381 ctggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     1441 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     1501 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1561 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     1621 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     1681 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     1741 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     1801 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     1861 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     1921 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     1981 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2041 aagatccttt gatcttttct acggggtctg acgctctatt caacaaagcc gccgtcccgt
     2101 caagtcagcg taaatgggta gggggcttca aatcgtcccc ccatacgata taagttgtta
     2161 ctagtgcttg gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa
     2221 tccagatgga gttctgaggt cattactgga tctatcaaca ggagtccaag cgagctcgat
     2281 atcaaattac gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct
     2341 gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac
     2401 cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat
     2461 attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa
     2521 catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc
     2581 ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga
     2641 aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac
     2701 cagctcaccg tctttcattg ccatacgaaa ttccggatga gcattcatca ggcgggcaag
     2761 aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc
     2821 cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc
     2881 aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt
     2941 ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag
     3001 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa cgtctcattg
     3061 atacctgaaa caaaacccat cgtacggcca aggaagtctc caataactgt gatccaccac
     3121 aagcgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtcat gcataatccg
     3181 cacgcatctg gaataaggaa gtgccattcc gcctgacct
//
