LOCUS Exported 7129 bp DNA circular SYN 17-JUN-2022 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7129) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 18, 2022 from SnapGene Viewer 6.0.2 https://www.snapgene.com FEATURES Location/Qualifiers source 1..7129 /mol_type="other DNA" /organism="synthetic DNA construct" 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" primer_bind 895..928 /label=Kpn1-BNP-pCDNA3-For CDS 2371..2382 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(2563..2596) /label=BamH1-BNP-V5-Rev primer_bind complement(2563..2596) /label=BamH1-BNP-stop-Rev CDS 2599..2640 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" promoter complement(2681..2699) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" polyA_signal 2725..2949 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2995..3423 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 3437..3767 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3618..3753 /label=SV40 ori /note="SV40 origin of replication" CDS 3834..4628 /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 4802..4923 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4972..4988) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 4996..5012 /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(5020..5050) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5065..5086 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(5374..5962) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6133..6993) /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(6994..7098) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttggtacc 901 gccaccatgt ccaacatgga tattgacggt ataaacactg ggacaattga caaaacaccg 961 gaagaaataa cttctggaac cagtgggaca accagaccaa tcattagacc agcaaccctt 1021 gccccaccaa gcaacaaacg aacccgtaac ccatccccgg aaagagcaac cacaagcagt 1081 gaagatgatg tcggaaggaa aacccaaaag aaacagaccc cgacagagat aaagaagagc 1141 gtctacaaca tggtggtgaa actgggcgaa ttctataacc agatgatggt caaagctgga 1201 ctcaatgatg acatggagag aaatctaatc caaaatgcgc atgccgtgga aagaattcta 1261 ttggctgcca ctgatgacaa gaaaaccgag ttccagaaga aaaagaatgc cagagatgtc 1321 aaagaaggga aagaagaaat agatcacaac aaaacaggag gcacctttta caagatggta 1381 agagatgata aaaccatcta cttcagccct ataagaatta cctttttaaa agaagaggtg 1441 aaaacaatgt acaaaaccac catggggagt gatggcttca gtggactaaa tcacataatg 1501 attgggcatt cacagatgaa tgatgtctgt ttccaaagat caaaggcact aaaaagagtt 1561 ggacttgatc cttcattaat cagtaccttt gcgggaagca cagtccccag aagatcaggt 1621 gcgactggtg ttgcaatcaa aggaggtgga accttagtgg ctgaagccat tcgatttata 1681 ggaagagcaa tggcagacag agggctattg agagacatca aagccaagac tgcctatgaa 1741 aagattcttc tgaatctaaa aaacaaatgc tctgcgcccc aacaaaaggc tctagttgat 1801 caagtgatcg gaagcagaaa tccggggatt gcagacattg aagatctaac cctgcttgct 1861 cgtagtatgg tcgttgttag gccctctgtg gcaagcaaag tggtgcttcc cataagcatt 1921 tacgccaaaa tacctcaact agggttcaat gttgaagagt actctatggt tgggtacgaa 1981 gccatggctc tttacaatat ggcaacacct gtgtccatat taagaatggg agatgatgca 2041 aaagataaat cgcaattatt cttcatgtct tgcttcggag ctgcctatga agacctgaga 2101 gttttgtctg cattaacagg cacagaattc aagcctagat cagcattaaa atgcaagggt 2161 ttccatgttc cagcaaagga acaggtagaa ggaatgggag cagctctgat gtccatcaag 2221 ctccagtttt gggctccgat gaccagatct ggggggaacg aagtaggtgg agacggaggg 2281 tctggccaaa taagctgcag cccagtgttt gcagtggaaa gacctattgc tctaagcaag 2341 caagctgtaa gaagaatgct gtcaatgaat attgagggac gtgatgcaga tgtcaaagga 2401 aatctactca agatgatgaa tgactcaatg gctaagaaaa ccagtggaaa tgctttcatt 2461 gggaagaaaa tgtttcaaat atcagacaaa aacaaaacca atcccattga aattccaatt 2521 aagcagacca tccccaattt cttctttggg agggacacag cagaggatta tgatgacctc 2581 gattatggtg gatccggagg taagcctatc cctaaccctc tcctcggtct cgattctacg 2641 tagtaagcgg ccgctcgagc atgcatctag agggccctat tctatagtgt cacctaaatg 2701 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc 2761 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa 2821 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg 2881 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg 2941 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc 3001 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac 3061 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg 3121 ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt 3181 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc 3241 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct 3301 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga 3361 ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga 3421 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccaggcag 3481 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag 3541 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc 3601 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc 3661 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat 3721 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag 3781 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg 3841 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg 3901 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg 3961 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg 4021 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg 4081 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc 4141 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc 4201 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc 4261 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc 4321 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg 4381 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct 4441 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt 4501 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc 4561 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt 4621 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc 4681 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg 4741 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc 4801 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 4861 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 4921 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct 4981 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat 5041 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc 5101 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 5161 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 5221 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 5281 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 5341 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 5401 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 5461 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 5521 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg 5581 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 5641 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 5701 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 5761 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt 5821 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 5881 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 5941 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 6001 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 6061 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 6121 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 6181 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 6241 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 6301 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 6361 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 6421 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 6481 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 6541 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 6601 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 6661 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 6721 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 6781 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 6841 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 6901 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 6961 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 7021 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 7081 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc //