LOCUS Exported 2710 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Level T acceptor vector for chromosomal integration with lacZ compatible with blue/white screening. Modified pUC19 vector.. ACCESSION . VERSION . KEYWORDS pCAT.015 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2710 /organism="synthetic DNA construct" /mol_type="other DNA" 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 141..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 908..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 //