LOCUS       sequence-361448-        3342 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  tdk/kanR dual selection cassette for engineering the genome of 
            Acinetobacter baylyi ADP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3342)
  AUTHORS   Suarez GA, Dugan KR, Renda BA, Leonard SP, Gangavarapu LS, Barrick 
            JE
  TITLE     Rapid and assured genetic engineering methods applied to 
            Acinetobacter baylyi ADP1 genome streamlining.
  JOURNAL   Nucleic Acids Res. 2020 May 7;48(8):4585-4600. doi: 
            10.1093/nar/gkaa204.
  PUBMED    32232367
REFERENCE   2  (bases 1 to 3342)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2020-05-7- 7"; volume: "48"; issue: "8";
            pages: "4585-4600"
COMMENT     Sequence Label: pBTK622
FEATURES             Location/Qualifiers
     source          1..3342
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             57..68
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(849..868)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             860..1675
                     /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             860..1675
                     /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(933..952)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      complement(1707..1741)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1812..2471)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2386..2405
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2653..3241
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3142..3161
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcggtctcaa acgcccagcc tccaattcaa atcataaaaa atttatttgc tttattaaag
       61 aggagaaatt aattaatggc acagctatat ttctactatt ccgcaatgaa tgcgggtaag
      121 tctacagcat tgttgcaatc ttcatacaat taccaggaac gcggcatgcg cactgtcgta
      181 tatacggcag aaattgatga tcgctttggt gccgggaaag tcagttcgcg tataggtttg
      241 tcatcgcctg caaaattatt taaccaaaat tcatcattat ttgatgagat tcgtgcggaa
      301 catgaacagc aggcaattca ttgcgtactg gttgatgaat gccagttttt aaccagacaa
      361 caagtatatg aattatcgga ggttgtcgat caactcgata tacccgtact ttgttatggt
      421 ttacgtaccg attttcgagg tgaattattt attggcagcc aatacttact ggcatggtcc
      481 gacaaactgg ttgaattaaa aaccatctgt ttttgtggcc gtaaagcaag catggtgctg
      541 cgtcttgatc aagcaggcag accttataac gaaggtgagc aggtggtaat tggtggtaat
      601 gaacgatacg tttctgtatg ccgtaaacac tataaagagg cgttacaagt cgactcatta
      661 acggctattc aggaaaggca tcgccacgat taagcggccg ccaccgcggt ggagctcggt
      721 acccggggat cctctagagc ggacccggga aagccacgtt gtgtctcaaa atctctgatg
      781 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
      841 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcttgct cgaggccgcg
      901 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
      961 gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct
     1021 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     1081 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     1141 atggttactc accactgcga tccccgggaa aacagcattc caggtattag aagaatatcc
     1201 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     1261 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     1321 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     1381 tgttgaacaa gtctggaaag aaatgcataa gcttttgcca ttctcaccgg attcagtcgt
     1441 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     1501 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     1561 ctgcctcggt gagttttctc cttcactaca gaaacggctt tttcaaaaat atggtattga
     1621 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaagcatg
     1681 cggagctggg ctgtgagacc agaccaataa aaaacgcccg gcggcaaccg agcgttctga
     1741 acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt ccaagcgagc
     1801 tcgatatcaa attacgcccc gccctgccac tcatcgcagt actgttgtaa ttcattaagc
     1861 attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc
     1921 agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc gaagaagttg
     1981 tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt ggctgaaacg
     2041 aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc gtaacacgcc
     2101 acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc actccagagc
     2161 gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac actatcccat
     2221 atcaccagct caccgtcttt cattgccata cgaaattccg gatgagcatt catcaggcgg
     2281 gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac ggtctttaaa
     2341 aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac tgactgaaat
     2401 gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata tccagtgatt
     2461 tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc
     2521 ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccc gatcaatcat
     2581 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     2641 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2701 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2761 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     2821 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2881 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2941 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3001 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3061 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3121 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3181 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3241 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3301 gttctttcct gcgttatccc ctgattctgt ggataaccgt ag
//
