LOCUS       sequence-250441-        3100 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level T acceptor vector for chromosomal integration with lacZ 
            compatible with blue/white screening. Modified pUC19 vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3100)
  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 3100)
  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.334
FEATURES             Location/Qualifiers
     source          1..3100
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(6..24)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        92..196
                     /gene="bla"
                     /label=AmpR promoter
     CDS             418..1209
                     /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"
     rep_origin      1618..2206
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2107..2126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2360..2377
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2494..2515
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2530..2535,2536..2553,2554..2560)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2568..2584
                     /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     2573..2595
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2592..2608
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2592..2608
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2604..2951
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLDALSSKTREDPRVPSSNSLAVVLQRRDWEN
                     PGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIW
                     CTLSTICSDAA"
     primer_bind     complement(2702..2719)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2702..2718)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2711..2733)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2927..2946)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3046..3068
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
       61 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
      121 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
      181 ttgaaaaagg aagagtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg
      241 tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttt tggggtacag
      301 tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta
      361 tggagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt aaacatcatg
      421 agggaagcgg tgatcgccga agtatcgact caactatcag aggtagttgg cgtcatcgag
      481 cgccatctcg aaccgacgtt gctggccgta catttgtacg gctccgcagt ggatggcggc
      541 ctgaagccac acagcgatat tgatttgctg gttacggtga ccgtaaggct tgatgaaaca
      601 acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg agagagcgag
      661 attctccgcg ctgtagaagt caccattgtt gtgcacgacg acatcattcc gtggcgttat
      721 ccagctaagc gcgaactgca atttggagaa tggcagcgca atgacattct tgcaggtatc
      781 ttcgagccag ccacgatcga cattgatctg gctatcttgc tgacaaaagc aagagaacat
      841 agcgttgcct tggtaggtcc agcggcggag gaactctttg atccggttcc tgaacaggat
      901 ctatttgagg cgctaaatga aaccttaacg ctatggaact cgccgcccga ctgggctggc
      961 gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa
     1021 atcgcgccga aggatgtcgc tgccgactgg gcaatggagc gcctgccggc ccagtatcag
     1081 cccgtcatac ttgaagctag acaggcttat cttggacaag aagaagatcg cttggcctcg
     1141 cgcgcagatc agttggaaga atttgtccat tacgtaaaag gcgagatcac caaggtagtc
     1201 ggcaaataac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     1261 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     1321 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcga cggggagtca
     1381 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     1441 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     1501 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     1561 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1621 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     1681 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     1741 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     1801 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     1861 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     1921 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     1981 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2041 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2101 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2161 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2221 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2281 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2341 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     2401 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     2461 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     2521 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     2581 acaatttcac acaggaaaca gctatgacca tgattacgcc aagcttgcat gcctgcaggt
     2641 cgactctaga tgccttgtct tcgaagacaa gggaggatcc ccgggtaccg agctcgaatt
     2701 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     2761 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     2821 gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc
     2881 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg
     2941 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3001 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3061 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
//
