LOCUS Exported 3761 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION pTDH2 in domestication vector. ACCESSION . VERSION . KEYWORDS pUPD-pTDH2 (01-11) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3761) AUTHORS Perez-Gonzalez A, Kniewel R, Veldhuizen M, Verma HK, Navarro-Rodriguez M, Rubio LM, Caro E TITLE Adaptation of the GoldenBraid modular cloning system and creation of a toolkit for the expression of heterologous proteins in yeast mitochondria. JOURNAL BMC Biotechnol. 2017 Nov 13;17(1):80. doi: 10.1186/s12896-017-0393-y. PUBMED 29132331 REFERENCE 2 (bases 1 to 3761) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3761 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..31 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 39..55 /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 44..66 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 63..79 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 63..79 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 97..115 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 97..114 /label=SP6 /note="SP6 promoter, forward primer" regulatory 955..964 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 959..968 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind complement(998..1017) /label=T7 /note="T7 promoter, forward primer" promoter complement(999..1017) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1024..1041) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(1024..1040) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1033..1055) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 1181..1636 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1268..1287) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1478..1499 /label=F1ori-F /note="F1 origin, forward primer" promoter 1714..1818 /gene="bla" /label=AmpR promoter CDS 1819..2679 /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(2037..2056) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2850..3438 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3339..3358 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3592..3609 /label=L4440 /note="L4440 vector, forward primer" protein_bind 3726..3747 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." ORIGIN 1 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca 61 cacaggaaac agctatgacc atgattacgc caagctattt aggtgacact atagaatact 121 caagctatgc atccaacgcg ttgggagctc tcccatatgg tcgacctgca ggcggccgcg 181 aattcactag tgattgggcg atgagtggtc tcgggagatt ggtttttcca gtgaatgatt 241 atttgtcgtt accctttcgt aaaagttcaa acacgttttt aagtattgtt tagttgctct 301 ttcgacatat atgattatcc ctgcgcggct aaagttaagg atgcaaaaaa cataagacaa 361 ctgaagttaa tttacgtcaa ttaagttttc cagggtaatg atgttttggg cttccactaa 421 ttcaataagt atgtcatgaa atacgttgtg aagagcatcc agaaataatg aaaagaaaca 481 acgaaactgg gtcggcctgt tgtttctttt ctttaccacg tgatctgcgg catttacagg 541 aagtcgcgcg ttttgcgcag ttgttgcaac gcagctacgg ctaacaaagc ctagtggaac 601 tcgactgatg tgttagggcc taaaactggt ggtgacagct gaagtgaact attcaatcca 661 atcatgtcat ggctgtcaca aagaccttgc ggaccgcacg tacgaacaca tacgtatgct 721 aatatgtgtt ttgatagtac ccagtgatcg cagacctgca atttttttgt aggtttggaa 781 gaatatataa aggttgcact cattcaagat agtttttttc ttgtgtgtct attcatttta 841 ttattgtttg tttaaatgtt aaaaaaacca agaacttagt ttcaaattaa attcatcaca 901 caaacaaaca aaacaaacca tcgagaccac tcatcgccca atcgaattcc cgcggccgcc 961 atggcggccg ggagcatgcg acgtcgggcc caattcgccc tatagtgagt cgtattacaa 1021 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa 1081 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga 1141 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca 1201 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 1261 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 1321 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 1381 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc cagcgccctg atagacggtt 1441 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 1501 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg 1561 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata 1621 ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac 1681 accgcatcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc 1741 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa 1801 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt 1861 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct 1921 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc 1981 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta 2041 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac 2101 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc 2161 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac 2221 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg 2281 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac 2341 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc 2401 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt 2461 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga 2521 gccggtgagc gtgggtcgcg cggtatcatt gcagcactgg ggccagatgg taagccctcc 2581 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag 2641 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca 2701 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc 2761 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca 2821 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc 2881 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta 2941 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt 3001 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc 3061 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg 3121 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg 3181 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag 3241 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc 3301 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat 3361 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg 3421 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc 3481 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt 3541 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca 3601 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg 3661 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac 3721 gcaattaatg tgagttagct cactcattag gcaccccagg c //