LOCUS       sequence-234015-        2710 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 2710)
  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 2710)
  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.015
FEATURES             Location/Qualifiers
     source          1..2710
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    105..126
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(141..146,147..164,165..171)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    179..195
                     /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     184..206
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     203..219
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     203..219
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             215..562
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLDALSSKTREDPRVPSSNSLAVVLQRRDWEN
                     PGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIW
                     CTLSTICSDAA"
     primer_bind     complement(313..330)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(313..329)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(322..344)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(538..557)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     657..679
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(717..735)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        803..907
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(908..976,977..1768)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1126..1145)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1939..2527
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2428..2447
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2681..2698
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
       61 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
      121 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
      181 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttgca
      241 tgcctgcagg tcgactctag atgccttgtc ttcgaagaca agggaggatc cccgggtacc
      301 gagctcgaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
      361 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
      421 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg
      481 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac
      541 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
      601 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
      661 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
      721 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
      781 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
      841 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
      901 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
      961 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     1021 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     1081 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     1141 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     1201 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     1261 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     1321 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     1381 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     1441 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     1501 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     1561 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     1621 tggttctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     1681 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     1741 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     1801 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     1861 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     1921 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     1981 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2041 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     2101 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2161 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2221 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2281 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2341 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2401 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2461 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2521 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     2581 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     2641 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     2701 agcggaagag
//
