LOCUS       sequence-259191-        3766 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3766)
  AUTHORS   Azubuike CC, Gatehouse AMR, Howard TP
  TITLE     pCAT vectors overcome inefficient electroporation of Cupriavidus 
            necator H16.
  JOURNAL   N Biotechnol. 2021 Nov 25;65:20-30. doi: 10.1016/j.nbt.2021.07.003. 
            Epub 2021 Jul 29.
  PUBMED    34333160
REFERENCE   2  (bases 1 to 3766)
  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: "N 
            Biotechnol."; date: "2021-11-25"; volume: "65:20-30. doi"; pages: " 
            10.1016/j.nbt.2021.07.003. Epub 2021 Jul 29"
COMMENT     Sequence Label: pCAT206
FEATURES             Location/Qualifiers
     source          1..3766
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(93..458)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWVPGPPSRSTVSISLISNSCSPGDPLV
                     LERPPPRWSSNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWKL"
     primer_bind     227..249
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     241..258
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     242..258
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     268..287
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        268..286
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    295..402
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     319..335
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     319..335
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(369..385)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(370..386)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(415..435)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(415..433)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(454..470)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(454..470)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(467..489)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    478..494
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(502..508,509..526,527..532))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     699..717
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(744..749,750..766,767..772)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             820..2010
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(1172..1191)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      2253..3022
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             3023..3685
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
ORIGIN
        1 gaacttaatt aactcatcgc agtcggccta ttggttaaaa aatgagctga tttaacaaaa
       61 atttaacgcg aattttaaca aaatattaac gcttacaatt tccattcgcc attcaggctg
      121 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
      181 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
      241 tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga attggagctc
      301 caccgcggtg gcggccgctc tagaactagt ggatcccccg ggctgcagga attcgatatc
      361 aagcttatcg ataccgtcga cctcgagggg gggcccggta cccagctttt gttcccttta
      421 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
      481 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
      541 tgcctaatga gtgaagcggc gcgccgttgg gaagccctgc aaagtaaact ggatggcttt
      601 cttgccgcca aggatctgat ggcgcagggg atcaagatct ccgaaaagtg ccacctgacg
      661 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
      721 ttcgtcttca agaattctca tgtttgacag cttatcatcg ataagcttta atgcggtagt
      781 ttatcacagt taaattgcta acgcagtcag gcaccgtgta tgaaatctaa caatgcgctc
      841 atcgtcatcc tcggcaccgt caccctggat gctgtaggca taggcttggt tatgccggta
      901 ctgccgggcc tcttgcggga tatcgtccat tccgacagca tcgccagtca ctatggcgtg
      961 ctgctagcgc tatatgcgtt gatgcaattt ctatgcgcac ccgttctcgg agcactgtcc
     1021 gaccgctttg gccgccgccc agtcctgctc gcttcgctac ttggagccac tatcgactac
     1081 gcgatcatgg cgaccacacc cgtcctgtgg atcctctacg ccggacgcat cgtggccggc
     1141 atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa
     1201 gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc
     1261 cccgtggccg ggggactgtt gggcgccatc tccttgcatg caccattcct tgcggcggcg
     1321 gtgctcaacg gcctcaacct actactgggc tgcttcctaa tgcaggagtc gcataaggga
     1381 gagcgtcgac cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
     1441 ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
     1501 gtgccggcag cgctctgggt cattttcggc gaggaccgct ttcgctggag cgcgacgatg
     1561 atcggcctgt cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc cttcgtcact
     1621 ggtcccgcca ccaaacgttt cggcgagaag caggccatta tcgccggcat ggcggccgac
     1681 gcgctgggct acgtcttgct ggcgttcgcg acgcgaggct ggatggcctt ccccattatg
     1741 attcttctcg cttccggcgg catcgggatg cccgcgttgc aggccatgct gtccaggcag
     1801 gtagatgacg accatcaggg acagcttcaa ggatcgctcg cggctcttac cagcctaact
     1861 tcgatcactg gaccgctgat cgtcacggcg atttatgccg cctcggcgag cacatggaac
     1921 gggttggcat ggattgtagg cgccgcccta taccttgtct gcctccccgc gttgcgtcgc
     1981 ggtgcatgga gccgggccac ctcgacctga atggaagccg gcggcacctc gctaacggat
     2041 tcaccactcc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     2101 ggattcatcg actgtggcag gtggccggcc cttcgcaaag tcgtgaccgc ctacggcggc
     2161 tgcggcgccc tacgggcttg ctctccgggc ttcgccctgc gcggtcgctg cgctcccttg
     2221 ccagcccgtg gatatgtgga cgatggccgc gagcggccac cggctggctc gcttcgctcg
     2281 gcccgtggac aaccctgctg gacaagctga tggacaggct gcgcctgccc acgagcttga
     2341 ccacagggat tgcccaccgg ctacccagcc ttcgaccaca tacccaccgg ctccaactgc
     2401 gcggcctgcg gccttgcccc atcaattttt ttaattttct ctggggaaaa gcctccggcc
     2461 tgcggcctgc gcgcttcgct tgccggttgg acaccaagtg gaaggcgggt caaggctcgc
     2521 gcagcgaccg cgcagcggct tggccttgac gcgcctggaa cgacccaagc ctatgcgagt
     2581 gggggcagtc gaaggcgaag cccgcccgcc tgccccccga gcctcacggc ggcgagtgcg
     2641 ggggttccaa gggggcagcg ccaccttggg caaggccgaa ggccgcgcag tcgatcaaca
     2701 agccccggag gggccacttt ttgccggagg gggagccgcg ccgaaggcgt gggggaaccc
     2761 cgcaggggtg cccttctttg ggcaccaaag aactagatat agggcgaaat gcgaaagact
     2821 taaaaatcaa caacttaaaa aaggggggta cgcaacagct cattgcggca ccccccgcaa
     2881 tagctcattg cgtaggttaa agaaaatctg taattgactg ccacttttac gcaacgcata
     2941 attgttgtcg cgctgccgaa aagttgcagc tgattgcgca tggtgccgca accgtgcggc
     3001 accctaccgc atggagataa gcatggccac gcagtccaga gaaatcggca ttcaagccaa
     3061 gaacaagccc ggtcactggg tgcaaacgga acgcaaagcg catgaggcgt gggccgggct
     3121 tattgcgagg aaacccacgg cggcaatgct gctgcatcac ctcgtggcgc agatgggcca
     3181 ccagaacgcc gtggtggtca gccagaagac actttccaag ctcatcggac gttctttgcg
     3241 gacggtccaa tacgcagtca aggacttggt ggccgagcgc tggatctccg tcgtgaagct
     3301 caacggcccc ggcaccgtgt cggcctacgt ggtcaatgac cgcgtggcgt ggggccagcc
     3361 ccgcgaccag ttgcgcctgt cggtgttcag tgccgccgtg gtggttgatc acgacgacca
     3421 ggacgaatcg ctgttggggc atggcgacct gcgccgcatc ccgaccctgt atccgggcga
     3481 gcagcaacta ccgaccggcc ccggcgagga gccgcccagc cagcccggca ttccgggcat
     3541 ggaaccagac ctgccagcct tgaccgaaac ggaggaatgg gaacggcgcg ggcagcagcg
     3601 cctgccgatg cccgatgagc cgtgttttct ggacgatggc gagccgttgg agccgccgac
     3661 acgggtcacg ctgccgcgcc ggtagcactt gggttgcgca gcaacccgta agtgcgctgt
     3721 tccagactat cggctgtagc cgcctcgccg ccctatacct tgtctg
//
