LOCUS Exported 2506 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Level 2 Destination vector - Position A - Assembly of 4-5 TU's from Level 1-ABCD or ABCDE. ACCESSION . VERSION . KEYWORDS pTU2-A-lacZ SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2506) AUTHORS Moore SJ, Lai HE, Kelwick RJ, Chee SM, Bell DJ, Polizzi KM, Freemont PS TITLE EcoFlex: A Multifunctional MoClo Kit for E. coli Synthetic Biology. JOURNAL ACS Synth Biol. 2016 May 2. PUBMED 27096716 REFERENCE 2 (bases 1 to 2506) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2506 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 36..52 /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 41..63 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 60..76 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 60..76 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" RBS 85..96 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" terminator 460..517 /label=his operon terminator /note="This putative transcriptin terminator from the E. coli his operon has a 2-bp deletion introduced during synthesis. Its efficiency has not been determined." primer_bind complement(541..558) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(712..1300) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(792..811) /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator 1483..1577 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(1598..2257) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind 2172..2191 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" primer_bind 2419..2437 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" terminator 2441..2484 /label=bacterial terminator /note="putative bacterial transcription terminator" misc_feature 2487..2 /label=BioBrick prefix /note="BioBrick prefix for parts that do not start with ""ATG""" ORIGIN 1 aggtctcact atatctagag acgtctagac atatgttgtg agcggataac aatttcacac 61 aggaaacagc tatgaccatg gtacaaagag gagaaaggac atgggtgacc cgaaactgct 121 gctggaagtt ccgcacgcta tctcttcttc tccgggtaac tctctggctg ttgttctgca 181 acgtcgtgac tgggaaaacc cgggtgttac ccagctgaac cgtctggctg ctcacccgcc 241 gttcgcttct tggcgtaact ctgaagaagc tcgtaccgac cgtccgtccc agcagctgcg 301 ttctctgatc cgtctgctga ccaaaccgga acgtaaactg tcttggctgc tgccgccgct 361 gtctaacaac taacgcaaaa aaccccgccc ctgacagggc ggggtttttt cgcactagtg 421 gatccgcatg ccgtctcatt aggtacagag accctgcagt ccggcaaaaa agggcaaggt 481 gtcaccaccc tgcccttttt ctttaaaacc gaaaagatta cttcgcgtta tgcaggcttc 541 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 601 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 661 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccacag 721 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 781 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 841 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 901 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 961 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 1021 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 1081 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 1141 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 1201 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt 1261 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 1321 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt 1381 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta 1441 aagtatatat gagtaaactt ggtctgacag ctcgaggctt ggattctcac caataaaaaa 1501 cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg 1561 atctatcaac aggagtccaa gcgagctcga tatcaaatta cgccccgccc tgccactcat 1621 cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat 1681 gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca 1741 tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga 1801 aactcaccca gggattggct gacacgaaaa acatattctc aataaaccct ttagggaaat 1861 aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga 1921 aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg 1981 tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgaa 2041 attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt 2101 gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat 2161 aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata 2221 tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa 2281 atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg 2341 aacctcttac gtgcccgatc aactcgagtg ccacctgacg tctaagaaac cattattatc 2401 atgacattaa cctataaaaa taggcgtatc acgaggcaga atttcagata aaaaaaatcc 2461 ttagctttcg ctaaggatga tttctggaat tcgcggccgc ttctag //