LOCUS Exported 7200 bp ds-DNA circular SYN 17-NOV-2016 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7200) AUTHORS . TITLE Direct Submission JOURNAL Exported Wednesday, Jun 19, 2019 from SnapGene Viewer 4.3.8 https://www.snapgene.com FEATURES Location/Qualifiers source 1..7200 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" gene 919..1527 /label=Rab8A TN gene 1552..2517 /label=BirA* CDS 2548..2574 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" polyA_signal 2800..3024 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3070..3498 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 3512..3841 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3692..3827 /label=SV40 ori /note="SV40 origin of replication" CDS 3908..4702 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" polyA_signal 4876..4997 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5046..5062) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 5070..5086 /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(5094..5124) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5139..5160 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(5448..6033) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6204..7064) /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" promoter complement(7065..7169) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 aagcttaccg gttgtaatat ggcgaagacc tacgattacc tgttcaagct gctgctgatc 961 ggggactcgg gggtggggaa gaactgtgtc ctgttccgct tctccgagga cgccttcaac 1021 tccactttta tctccaccat aggaattgac tttaaaatta ggaccataga gctcgatggc 1081 aagagaatta aactgcagat atgggacaca gccggtcagg aacggtttcg gacgatcaca 1141 acggcctact acaggggtgc aatgggcatc atgctggtct acgacatcac caacgagaag 1201 tccttcgaca acatccggaa ctggattcgc aacattgagg agcacgcctc tgcagacgtc 1261 gaaaagatga tactcgggaa caagtgtgat gtgaatgaca agagacaagt ttccaaggaa 1321 cggggagaaa agctggccct cgactatgga atcaagttca tggagaccag cgcgaaggcc 1381 aacatcaatg tggaaaatgc atttttcact ctcgccagag atatcaaagc aaaaatggac 1441 aaaaaattgg aaggcaacag cccccagggg agcaaccagg gagtcaaaat cacaccggac 1501 cagcagaaga ggagcagctt tttccgagaa ttcggagccg gagctggtac catggacaag 1561 gacaacaccg tgcccctgaa gctgatcgcc ctgctggcca acggcgagtt ccactctggc 1621 gagcagctgg gagagaccct gggaatgagc agagccgcca tcaacaagca catccagaca 1681 ctgagagact ggggagtgga cgtgttcacc gtgcctggca agggctacag cctgcctgag 1741 cctatccagc tgctgaacgc caagcagatc ctgggacagc tggatggcgg aagcgtggcc 1801 gtgctgcctg tgatcgactc caccaatcag tacctgctgg acagaatcgg agagctgaag 1861 tccggcgacg cctgcatcgc cgagtaccag caggctggca gaggaggcag aggacggaag 1921 tggttcagcc cattcggagc caacctgtac ctgtccatgt tctggagact ggagcaggga 1981 cctgctgctg ccatcggact gagtctggtg atcggaatcg tgatggccga ggtgctgaga 2041 aagctgggag ccgacaaggt gagagtgaag tggcctaatg acctgtacct ccaggaccgc 2101 aagctggctg gcatcctggt ggagctgaca ggcaagacag gcgatgccgc tcagatcgtg 2161 atcggagccg gaatcaacat ggccatgaga agagtggagg agagcgtggt gaaccagggc 2221 tggatcaccc tgcaggaggc tggcatcaac ctggaccgga acaccctggc cgccatgctg 2281 atcagagagc tgagagccgc tctggagctg ttcgagcagg agggactggc tccttacctg 2341 agcagatggg agaagctgga caacttcatc aacagacctg tgaagctgat catcggcgac 2401 aaggaaatct tcggcatctc cagaggaatc gacaagcagg gagctctgct gctggagcag 2461 gacggaatca tcaagccctg gatgggcgga gaaatctccc tgagaagcgc agagaaggga 2521 tccgtcgacg gagccggtgc tggcgcatac ccctacgacg tgcccgacta cgcaggctcc 2581 gcagttccag cggtagaaat cacccacacc ttctgggaaa ggaacctgcc ctctgtttct 2641 ggcctgctga agatcattgg attttccaca tcagtaactg ccctggggtt tgtgctgtac 2701 aaatacaagc tcctgccacg gtcttgaccc ggggatatcc agcacagtgg cggccgctcg 2761 agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc 2821 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg 2881 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc 2941 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg 3001 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg 3061 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca 3121 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct 3181 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt 3241 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac 3301 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct 3361 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt 3421 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac 3481 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc 3541 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg 3601 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc 3661 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc 3721 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc 3781 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa 3841 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc 3901 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag 3961 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg 4021 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa 4081 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc 4141 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc 4201 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga 4261 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa 4321 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct 4381 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat 4441 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt 4501 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta 4561 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga 4621 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg 4681 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg 4741 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc 4801 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag 4861 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc 4921 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa 4981 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa 5041 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 5101 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 5161 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 5221 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg 5281 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 5341 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 5401 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 5461 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 5521 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 5581 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 5641 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 5701 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 5761 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 5821 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 5881 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 5941 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag 6001 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg 6061 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa 6121 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata 6181 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg 6241 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata 6301 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg 6361 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct 6421 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt 6481 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc 6541 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga 6601 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt 6661 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc 6721 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa 6781 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca 6841 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca 6901 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct 6961 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc 7021 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa 7081 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt 7141 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc //