LOCUS       sequence-234003-        4355 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Self replication Level T acceptor vector with LacZ compatible 
            blue/white screening. Modified SEVA vector, RK2 ori vector. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4355)
  AUTHORS   Vasudevan R, Gale GAR, Schiavon AA, Puzorjov A, Malin J, Gillespie 
            MD, Vavitsas K, Zulkower V, Wang B, Howe CJ, Lea-Smith DJ, McCormick
            AJ
  TITLE     CyanoGate: A Modular Cloning Suite for Engineering Cyanobacteria 
            Based on the Plant MoClo Syntax.
  JOURNAL   Plant Physiol. 2019 May;180(1):39-55. doi: 10.1104/pp.18.01401. Epub
            2019 Feb 28.
  PUBMED    30819783
REFERENCE   2  (bases 1 to 4355)
  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: 
            "10.1104/pp.18.01401"; journalName: "Plant Physiol"; date: 
            "2019-05"; volume: "180"; issue: "1"; pages: "39-55"
COMMENT     Sequence Label: pCAT.011
FEATURES             Location/Qualifiers
     source          1..4355
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    121..142
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(157..162,163..180,181..187)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    195..211
                     /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)."
     primer_bind     200..222
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     219..235
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     219..235
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             231..554
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    248..304
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(305..322)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(305..321)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(314..336)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(530..549)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     terminator      612..706
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             942..1733
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     oriT            1879..1987
                     /note="incP origin of transfer"
     CDS             complement(2048..3196)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      3589..4120
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(4247..4333)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaagaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
       61 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
      121 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
      181 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
      241 ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccgggta ccgagctcga
      301 attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
      361 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
      421 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc
      481 tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct
      541 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgaa ctagtcttgg
      601 aagactcggg agactcctgt tgatagatcc agtaatgacc tcagaactcc atctggattt
      661 gttcagaacg ctcggttgcc gccgggcgtt ttttattggt gagaatccag gggtccccaa
      721 taattacgat ttacgtattt aaatgaacct tgaccgaacg cagcggtggt aacggcgcag
      781 tggcggtttt catggcttgt tatgactgtt tttttggggt acagtctatg cctcgggcat
      841 ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc agcaacgatg
      901 ttacgcagca gggcagtcgc cctaaaacaa agttaaacat catgagggaa gcggtgatcg
      961 ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga
     1021 cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagtg
     1081 atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga
     1141 tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag
     1201 aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac
     1261 tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga
     1321 tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag
     1381 gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa
     1441 atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag
     1501 tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg
     1561 tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag
     1621 ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca gatcagttgg
     1681 aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa taagacaatt
     1741 gtccttttcc gctgcataac cctgcttcgg ggtcattata gcgatttttt cggtatatcc
     1801 atcctttttc gcacgatata caggattttg ccaaagggtt cgtgtagact ttccttggtg
     1861 tatccaacgg cgtcagccgg gcaggatagg tgaagtaggc ccacccgcga gcgggtgttc
     1921 cttcttcact gtcccttatt cgcacctggc ggtgctcaac gggaatcctg ctctgcgagg
     1981 ctggccgtag gccggccgcg atgcaggtgg ctgctgaacc cccagccgga actgacccca
     2041 caaggcccta gcgtttgcaa tgcaccaggt catcattgac ccaggcgtgt tccaccaggc
     2101 cgctgcctcg caactcttcg caggcttcgc cgacctgctc gcgccacttc ttcacgcggg
     2161 tggaatccga tccgcacatg aggcggaagg tttccagctt gagcgggtac ggctcccggt
     2221 gcgagctgaa atagtcgaac atccgtcggg ccgtcggcga cagcttgcgg tacttctccc
     2281 atatgaattt cgtgtagtgg tcgccagcaa acagcacgac gatttcctcg tcgatcagga
     2341 cctggcaacg ggacgttttc ttgccacggt ccaggacgcg gaagcggtgc agcagcgaca
     2401 ccgattccag gtgcccaacg cggtcggacg tgaagcccat cgccgtcgcc tgtaggcgcg
     2461 acaggcattc ctcggccttc gtgtaatacc ggccattgat cgaccagccc aggtcctggc
     2521 aaagctcgta gaacgtgaag gtgatcggct cgccgatagg ggtgcgcttc gcgtactcca
     2581 acacctgctg ccacaccagt tcgtcatcgt cggcccgcag ctcgacgccg gtgtaggtga
     2641 tcttcacgtc cttgttgacg tggaaaatga ccttgttttg cagcgcctcg cgcgggattt
     2701 tcttgttgcg cgtggtgaac agggcagagc gggccgtgtc gtttggcatc gctcgcatcg
     2761 tgtccggcca cggcgcaata tcgaacaagg aaagctgcat ttccttgatc tgctgcttcg
     2821 tgtgtttcag caacgcggcc tgcttggctt cgctgacctg ttttgccagg tcctcgccgg
     2881 cggtttttcg cttcttggtc gtcatagttc ctcgcgtgtc gatggtcatc gacttcgcca
     2941 aacctgccgc ctcctgttcg agacgacgcg aacgctccac ggcggccgat ggcgcgggca
     3001 gggcaggggg agccagttgc acgctgtcgc gctcgatctt ggccgtagct tgctggacta
     3061 tcgagccgac ggactggaag gtttcgcggg gcgcacgcat gacggtgcgg cttgcgatgg
     3121 tttcggcatc ctcggcggaa aaccccgcgt cgatcagttc ttgcctgtat gccttccggt
     3181 caaacgtccg attcattcac cctccttgcg ggattgcccc ggaattaatt ccccggatcg
     3241 atccgtcgat cttgatcccc tgcgccatca gatccttggc ggcaagaaag ccatccagtt
     3301 tactttgcag ggcttcccaa ccttaccaga gggcgcccca gctggcaatt ccggttcgct
     3361 tgctgtccat aaaaccgccc agtctagcta tcgccatgta agcccactgc aagctacctg
     3421 ctttctcttt gcgcttgcgt tttcccttgt ccagatagcc cagtagctga cattcatccg
     3481 gggtcagcac cgtttctgcg gactggcttt ctacgtggct gccatttttg gggtgaggcc
     3541 gttcgcggcc gaggggcgca gcccctgggg ggatgggagg cccgcgttag cgggccggga
     3601 gggttcgaga agggggggca ccccccttcg gcgtgcgcgg tcacgcgcac agggcgcagc
     3661 cctggttaaa aacaaggttt ataaatattg gtttaaaagc aggttaaaag acaggttagc
     3721 ggtggccgaa aaacgggcgg aaacccttgc aaatgctgga ttttctgcct gtggacagcc
     3781 cctcaaatgt caataggtgc gcccctcatc tgtcagcact ctgcccctca agtgtcaagg
     3841 atcgcgcccc tcatctgtca gtagtcgcgc ccctcaagtg tcaataccgc agggcactta
     3901 tccccaggct tgtccacatc atctgtggga aactcgcgta aaatcaggcg ttttcgccga
     3961 tttgcgaggc tggccagctc cacgtcgccg gccgaaatcg agcctgcccc tcatctgtca
     4021 acgccgcgcc gggtgagtcg gcccctcaag tgtcaacgtc cgcccctcat ctgtcagtga
     4081 gggccaagtt ttccgcgagg tatccacaac gccggcggcc ctacatggct ctgctgtagt
     4141 gagtgggttg cgctccggca gcggtcctga tcccccgcag aaaaaaagga tctcaagaag
     4201 atcctttgat cttttctacg gcgcgcccag ctgtctaggg cggcggattt gtcctactca
     4261 ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtctttc gactgagcct
     4321 ttcgttttat ttgatgcctt gccgagtctt cttaa
//
