LOCUS       sequence-402926-        3325 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 3325)
  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 3325)
  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_44 [hph]
FEATURES             Location/Qualifiers
     source          1..3325
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             82..1107
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             82..1107
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     primer_bind     complement(1369..1385)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1369..1385)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1382..1404)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1557..2145)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1637..1656)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2278..2372)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2393..3052)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2967..2986
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3053..3155)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     3242..3264
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3256..3273
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3257..3273
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tagcgtgcga acagcacgct attgttgtat cattactcgc atccattctc aggctgtctc
       61 gtctcgtctc cgaggtctca aatgaaaaag cctgaactca ccgcgacgtc tgtcgagaag
      121 tttctgatcg aaaagttcga cagcgtgtcc gacctgatgc agctctcgga gggcgaagaa
      181 tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc
      241 gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg
      301 attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc
      361 cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag
      421 ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc
      481 ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg
      541 attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc
      601 gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac
      661 ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg
      721 gtcattgact ggagcgaggc aatgttcggg gattcccaat acgaggtcgc caacatcttc
      781 ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat
      841 ccggagcttg caggatcgcc acgactccgg gcgtatatgc tccgcattgg tcttgaccaa
      901 ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc
      961 gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc
     1021 gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc
     1081 agcactcgtc cgagggcaaa ggaatgagct ttgagaccac cccaggatac atagattacc
     1141 acaactccga gcccttccac ctagcgtgct aaccacgcac gctattgttg tatcccatat
     1201 aatgcaatcg tagtctacct gactgtactt agaaatgtgg cttcgccttt gcccacgcac
     1261 ctgatcgctc ctcgtttgct tttaaggacc ggacgaacca cagagcatta gaaggctagg
     1321 tggaggctca gtgatgataa gtctgcgatg gtggatgcat gtgtcatggt catagctgtt
     1381 tcctgtgtga aattgttatc cgctcagagg gcacaatcct attccgcgct atccgacaat
     1441 ctccaagaca ttaggtggag ttcagttcgg cgtatggcat atgtcgctgg aaagaacatg
     1501 tgagcaaaag gcctgcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     1561 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     1621 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     1681 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     1741 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     1801 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     1861 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     1921 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     1981 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     2041 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     2101 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     2161 ttttctacgg ggtctgacgc tctattcaac aaagccgccg tcccgtcaag tcagcgtaaa
     2221 tgggtagggg gcttcaaatc gtccccccat acgatataag ttgttactag tgcttggatt
     2281 ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca gatggagttc
     2341 tgaggtcatt actggatcta tcaacaggag tccaagcgag ctcgatatca aattacgccc
     2401 cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc
     2461 catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg
     2521 tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta
     2581 aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata ttctcaataa
     2641 accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt
     2701 gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt
     2761 gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt
     2821 tcattgccat acgaaattcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg
     2881 ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct
     2941 gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac
     3001 gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt
     3061 ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat
     3121 tatggtgaaa gttggaacct cttacgtgcc gatcaacgtc tcattgatac ctgaaacaaa
     3181 acccatcgta cggccaagga agtctccaat aactgtgatc caccacaagc gccagggttt
     3241 tcccagtcac gacgttgtaa aacgacggcc agtcatgcat aatccgcacg catctggaat
     3301 aaggaagtgc cattccgcct gacct
//
