LOCUS Exported 2606 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION MoClo Destination Vector: (based on pSB1A2 with fixed illegal site) [E:LacZa:H]. ACCESSION . VERSION . KEYWORDS DVA_EH SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2606) AUTHORS Iverson SV, Haddock TL, Beal J, Densmore DM TITLE CIDAR MoClo: Improved MoClo Assembly Standard and New E. coli Part Library Enable Rapid Combinatorial Design for Synthetic and Traditional Biology. JOURNAL ACS Synth Biol. 2016 Jan 15;5(1):99-103. doi: 10.1021/acssynbio.5b00124. Epub 2015 Nov 4. PUBMED 26479688 REFERENCE 2 (bases 1 to 2606) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2606 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 199..221 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 213..230 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 214..230 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" misc_feature 231..287 /label=MCS /note="pUC18/19 multiple cloning site" primer_bind complement(300..316) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(300..316) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(313..335) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 324..340 /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(348..378) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 393..414 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." misc_feature 514..534 /label=BioBrick suffix /note="universal suffix for all parts" primer_bind complement(540..557) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(711..1299) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(791..810) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1470..2330) /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 2093..2112 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2331..2435) /gene="bla" /label=AmpR promoter primer_bind 2503..2521 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" terminator 2525..2568 /label=bacterial terminator /note="putative bacterial transcription terminator" misc_feature 2571..2592 /label=BioBrick prefix /note="BioBrick prefix for parts that do not start with ""ATG""" CDS complement(2587..304) /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCRRHKLRP TSL" ORIGIN 1 gcttatgtct tctgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat 61 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc 121 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt 181 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcgagc 241 tcggtacccg gggatcctct agagtcgacc tgcaggcatg caagcttggc gtaatcatgg 301 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc 361 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg 421 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc 481 ggccaacgcg cggggaagac gtactaagag acctactagt agcggccgct gcaggcttcc 541 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 601 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 661 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccacagg 721 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 781 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 841 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 901 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 961 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 1021 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt 1081 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc 1141 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa 1201 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt 1261 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 1321 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 1381 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 1441 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 1501 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 1561 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg cgacccacgc 1621 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 1681 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 1741 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg 1801 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt 1861 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc 1921 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt 1981 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc 2041 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc 2101 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa 2161 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac 2221 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa 2281 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt 2341 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa 2401 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct 2461 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg 2521 cagaatttca gataaaaaaa atccttagct ttcgctaagg atgatttctg gaattcgcgg 2581 ccgcttctag agactagtgg gtctca //