LOCUS       sequence-401224-        3094 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 3094)
  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 3094)
  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_46 [nptII]
FEATURES             Location/Qualifiers
     source          1..3094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             82..876
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(136..155)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     746..765
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1138..1154)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1138..1154)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1151..1173)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1326..1914)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1406..1425)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2047..2141)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2162..2821)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2736..2755
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2822..2924)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     3011..3033
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3025..3042
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3026..3042
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tagcgtgcga acagcacgct attgttgtat cattactcgc atccattctc aggctgtctc
       61 gtctcgtctc cgaggtctca aatgattgaa caagatggat tgcacgcagg ttctccggcc
      121 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
      181 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
      241 tccggtgccc tgaatgaact ccaagacgag gcagcgcggc tatcgtggct ggccacgacg
      301 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
      361 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
      421 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
      481 gaccaccaag cgaaacaccg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
      541 gatcaggatg atctggacga agaacatcag gggctcgcgc cagccgaact gttcgccagg
      601 ctcaaggcgc ggatgcccga cggcgaggat ctcgtcgtga cccacggcga cgcctgcttg
      661 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
      721 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agaacttggc
      781 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
      841 atagccttct atcgccttct tgacgagttc ttctgagctt tgagaccacc ccaggataca
      901 tagattacca caactccgag cccttccacc tagcgtgcta accacgcacg ctattgttgt
      961 atcccatata atgcaatcgt agtctacctg actgtactta gaaatgtggc ttcgcctttg
     1021 cccacgcacc tgatcgctcc tcgtttgctt ttaaggaccg gacgaaccac agagcattag
     1081 aaggctaggt ggaggctcag tgatgataag tctgcgatgg tggatgcatg tgtcatggtc
     1141 atagctgttt cctgtgtgaa attgttatcc gctcagaggg cacaatccta ttccgcgcta
     1201 tccgacaatc tccaagacat taggtggagt tcagttcggc gtatggcata tgtcgctgga
     1261 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     1321 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     1381 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     1441 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     1501 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     1561 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     1621 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     1681 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     1741 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     1801 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     1861 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     1921 cctttgatct tttctacggg gtctgacgct ctattcaaca aagccgccgt cccgtcaagt
     1981 cagcgtaaat gggtaggggg cttcaaatcg tccccccata cgatataagt tgttactagt
     2041 gcttggattc tcaccaataa aaaacgcccg gcggcaaccg agcgttctga acaaatccag
     2101 atggagttct gaggtcatta ctggatctat caacaggagt ccaagcgagc tcgatatcaa
     2161 attacgcccc gccctgccac tcatcgcagt actgttgtaa ttcattaagc attctgccga
     2221 catggaagcc atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt
     2281 cgccttgcgt ataatatttg cccatggtga aaacgggggc gaagaagttg tccatattgg
     2341 ccacgtttaa atcaaaactg gtgaaactca cccagggatt ggctgagacg aaaaacatat
     2401 tctcaataaa ccctttaggg aaataggcca ggttttcacc gtaacacgcc acatcttgcg
     2461 aatatatgtg tagaaactgc cggaaatcgt cgtggtattc actccagagc gatgaaaacg
     2521 tttcagtttg ctcatggaaa acggtgtaac aagggtgaac actatcccat atcaccagct
     2581 caccgtcttt cattgccata cgaaattccg gatgagcatt catcaggcgg gcaagaatgt
     2641 gaataaaggc cggataaaac ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa
     2701 tatccagctg aacggtctgg ttataggtac attgagcaac tgactgaaat gcctcaaaat
     2761 gttctttacg atgccattgg gatatatcaa cggtggtata tccagtgatt tttttctcca
     2821 ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc
     2881 ttatttcatt atggtgaaag ttggaacctc ttacgtgccg atcaacgtct cattgatacc
     2941 tgaaacaaaa cccatcgtac ggccaaggaa gtctccaata actgtgatcc accacaagcg
     3001 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtcatgcata atccgcacgc
     3061 atctggaata aggaagtgcc attccgcctg acct
//
