LOCUS       sequence-401267-        3103 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 3103)
  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 3103)
  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_42 [aac3-IV]
FEATURES             Location/Qualifiers
     source          1..3103
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             82..882
                     /codon_start=1
                     /gene="aac(3)IV"
                     /product="aminoglycoside 3-N-acetyltransferase type IV     
                               "
                     /label=ApmR
                     /note="confers resistance to apramycin (Paget and Davies, 
                     1996)"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     primer_bind     complement(1147..1163)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1147..1163)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1160..1182)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1335..1923)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1415..1434)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2056..2150)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2171..2830)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2745..2764
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2831..2933)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     3020..3042
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3034..3051
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3035..3051
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tagcgtgcga acagcacgct attgttgtat cattactcgc atccattctc aggctgtctc
       61 gtctcgtctc cgaggtctca aatgtcatca gcggtggagt gcaatgtcgt gcaatacgaa
      121 tggcgaaaag ccgagctgat cggtcagctt ctcaaccttg gggttacccc cggcggtgtg
      181 ctgctggtcc acagctcctt ccgtagcgtc cggcccctcg aagatgggcc acttggactg
      241 atcgaggccc tgcgtgctgc gctgggtccg ggagggacgc tcgtgatgcc ctcgtggtca
      301 ggtctggacg acgagccgtt cgatcctgcc acgtcgcccg ttacaccgga ccttggagtt
      361 gtctctgaca cattctggcg cctgccaaat gtaaagcgca gcgcccatcc atttgccttt
      421 gcggcagcgg ggccacaggc agagcagatc atctctgatc cattgcccct gccacctcac
      481 tcgcctgcaa gcccggtcgc ccgtgtccat gaactcgatg ggcaggtact tctcctcggc
      541 gtgggacacg atgccaacac gacgctgcat cttgccgagt tgatggcaaa ggttccctat
      601 ggggtgccga gacactgcac cattcttcag gatggcaagt tggttcgcgt cgattatctc
      661 gagaatgacc actgctgtga gcgctttgcc ttggcggaca ggtggctcaa ggagaaaagc
      721 cttcagaagg aaggtccagt cggtcatgcc tttgctcggt tgatccgctc ccgcgacatt
      781 gtggcgacag ccctgggtca actgggccga gatccgttga tcttcctgca tccgccagag
      841 gcgggatgcg aagaatgcga tgccgctcgc cagtcgattg gctgagcttt gagaccaccc
      901 caggatacat agattaccac aactccgagc ccttccacct agcgtgctaa ccacgcacgc
      961 tattgttgta tcccatataa tgcaatcgta gtctacctga ctgtacttag aaatgtggct
     1021 tcgcctttgc ccacgcacct gatcgctcct cgtttgcttt taaggaccgg acgaaccaca
     1081 gagcattaga aggctaggtg gaggctcagt gatgataagt ctgcgatggt ggatgcatgt
     1141 gtcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcagagggc acaatcctat
     1201 tccgcgctat ccgacaatct ccaagacatt aggtggagtt cagttcggcg tatggcatat
     1261 gtcgctggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     1321 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     1381 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     1441 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     1501 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     1561 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     1621 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     1681 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     1741 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     1801 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     1861 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     1921 caagaagatc ctttgatctt ttctacgggg tctgacgctc tattcaacaa agccgccgtc
     1981 ccgtcaagtc agcgtaaatg ggtagggggc ttcaaatcgt ccccccatac gatataagtt
     2041 gttactagtg cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa
     2101 caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct
     2161 cgatatcaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca
     2221 ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc agcggcatca
     2281 gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt
     2341 ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgagacga
     2401 aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca
     2461 catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg
     2521 atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata
     2581 tcaccagctc accgtctttc attgccatac gaaattccgg atgagcattc atcaggcggg
     2641 caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa
     2701 aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg
     2761 cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt
     2821 ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg
     2881 gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaacgtctc
     2941 attgatacct gaaacaaaac ccatcgtacg gccaaggaag tctccaataa ctgtgatcca
     3001 ccacaagcgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tcatgcataa
     3061 tccgcacgca tctggaataa ggaagtgcca ttccgcctga cct
//
