LOCUS       sequence-401217-        3115 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 3115)
  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 3115)
  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_29 [aph(3')-I EZ-Tn5]
FEATURES             Location/Qualifiers
     source          1..3115
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             82..897
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             82..897
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(155..174)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     complement(1159..1175)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1159..1175)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1172..1194)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1347..1935)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1427..1446)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2068..2162)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2183..2842)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2757..2776
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2843..2945)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     3032..3054
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3046..3063
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3047..3063
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tagcgtgcga acagcacgct attgttgtat cattactcgc atccattctc aggctgtctc
       61 gtctcgtctc cgaggtctca aatgagccat attcaacggg aaacgtcttg ctcgaggccg
      121 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
      181 gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt
      241 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
      301 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
      361 gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat
      421 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
      481 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
      541 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
      601 cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc
      661 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
      721 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
      781 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
      841 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaagct
      901 ttgagaccac cccaggatac atagattacc acaactccga gcccttccac ctagcgtgct
      961 aaccacgcac gctattgttg tatcccatat aatgcaatcg tagtctacct gactgtactt
     1021 agaaatgtgg cttcgccttt gcccacgcac ctgatcgctc ctcgtttgct tttaaggacc
     1081 ggacgaacca cagagcatta gaaggctagg tggaggctca gtgatgataa gtctgcgatg
     1141 gtggatgcat gtgtcatggt catagctgtt tcctgtgtga aattgttatc cgctcagagg
     1201 gcacaatcct attccgcgct atccgacaat ctccaagaca ttaggtggag ttcagttcgg
     1261 cgtatggcat atgtcgctgg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     1321 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     1381 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1441 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     1501 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     1561 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     1621 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     1681 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     1741 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     1801 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     1861 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     1921 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tctattcaac
     1981 aaagccgccg tcccgtcaag tcagcgtaaa tgggtagggg gcttcaaatc gtccccccat
     2041 acgatataag ttgttactag tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc
     2101 gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag
     2161 tccaagcgag ctcgatatca aattacgccc cgccctgcca ctcatcgcag tactgttgta
     2221 attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg
     2281 ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg
     2341 cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat
     2401 tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac
     2461 cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt
     2521 cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa
     2581 cactatccca tatcaccagc tcaccgtctt tcattgccat acgaaattcc ggatgagcat
     2641 tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta
     2701 cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa
     2761 ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat
     2821 atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa
     2881 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     2941 gatcaacgtc tcattgatac ctgaaacaaa acccatcgta cggccaagga agtctccaat
     3001 aactgtgatc caccacaagc gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     3061 agtcatgcat aatccgcacg catctggaat aaggaagtgc cattccgcct gacct
//
