LOCUS V040524 6697 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040524 VERSION V040524 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 235..614 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 615..818 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 863..881 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 928..2133 /label="OPN3(NM_014322)" /note="OPN3(NM_014322)" /gene="OPN3" CDS 2140..2163 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" CDS 2170..2259 /codon_start=1 /product="two Strep-Tag®II moieties connected by a linker for enhanced binding to Strep-Tactin®, an engineered form of streptavidin (Schmidt et al., 2013)" /transl_table=1 /translation="SA,WSHPQFEK,GGGSGGGSGGSA,WSHPQFEK" /label="Twin-Strep-tag" polyA_signal 2297..2521 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2567..2995 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3009..3338 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3405..4199 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 4373..4494 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4543..4559) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4567..4583 /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-β-D-thiogalactopyranoside (IPTG)." /label="lac operator" promoter complement(4591..4621) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4636..4657 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4945..5530) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5701..6561) /codon_start=1 /gene="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /product="β-lactamase" /transl_table=1 /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA SLIKHW*" /label="AmpR" promoter complement(6562..6666) /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 gtttaaactt aagctgaatt cgccaccatg tactcgggga accgcagcgg cggccacggc 961 tactgggacg gcggcggggc cgcgggcgct gaggggccgg cgccggcggg gacactgagc 1021 cccgcgcccc tcttcagccc cggcacctac gagcgcctgg cgctgctgct gggctccatt 1081 gggctgctgg gcgtcggcaa caacctgctg gtgctcgtcc tctactacaa gttccagcgg 1141 ctccgcactc ccactcacct cctcctggtc aacatcagcc tcagcgacct gctggtgtcc 1201 ctcttcgggg tcacctttac cttcgtgtcc tgcctgagga acggctgggt gtgggacacc 1261 gtgggctgcg tgtgggacgg gtttagcggc agcctcttcg ggattgtttc cattgccacc 1321 ctaaccgtgc tggcctatga acgttacatt cgcgtggtcc atgccagagt gatcaatttt 1381 tcctgggcct ggagggccat tacctacatc tggctctact cactggcgtg ggcaggagca 1441 cctctcctgg gatggaacag gtacatcctg gacgtacacg gactaggctg cactgtggac 1501 tggaaatcca aggatgccaa cgattcctcc tttgtgcttt tcttatttct tggctgcctg 1561 gtggtgcccc tgggtgtcat agcccattgc tatggccata ttctatattc cattcgaatg 1621 cttcgttgtg tggaagatct tcagacaatt caagtgatca agattttaaa atatgaaaag 1681 aaactggcca aaatgtgctt tttaatgata ttcaccttcc tggtctgttg gatgccttat 1741 atcgtgatct gcttcttggt ggttaatggt catggtcacc tggtcactcc aacaatatct 1801 attgtttcgt acctctttgc taaatcgaac actgtataca atccagtgat ttatgtcttc 1861 atgatcagaa agtttcgaag atcccttttg cagcttctgt gcctccgact gctgaggtgc 1921 cagaggcctg ctaaagacct accagcagct ggaagtgaaa tgcagatcag acccattgtg 1981 atgtcacaga aagatgggga caggccaaag aaaaaagtga ctttcaactc ttcttccatc 2041 atttttatca tcaccagtga tgaatcactg tcagttgacg acagcgacaa aaccaatggg 2101 tccaaagttg atgtaatcca agttcgtcct ttgctcgagg attacaagga tgacgacgat 2161 aagggatcca gcgcctggtc ccaccctcag tttgagaagg gcggaggctc tggcggcgga 2221 agcggaggat ctgcttggag ccacccccag ttcgaaaagt gatgagggcc cgtttaaacc 2281 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc 2341 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa 2401 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac 2461 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg 2521 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc 2581 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc 2641 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt 2701 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac 2761 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag 2821 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa 2881 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg 2941 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc 3001 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta 3061 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag 3121 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa 3181 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 3241 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt 3301 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat 3361 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg 3421 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac 3481 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg 3541 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc 3601 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg 3661 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc 3721 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc 3781 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta 3841 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg 3901 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg 3961 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat 4021 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc 4081 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta 4141 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag 4201 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt 4261 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg 4321 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt 4381 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 4441 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 4501 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt 4561 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa 4621 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc 4681 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag 4741 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 4801 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 4861 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 4921 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 4981 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 5041 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 5101 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 5161 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 5221 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 5281 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 5341 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat 5401 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 5461 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 5521 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 5581 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 5641 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 5701 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 5761 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 5821 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 5881 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 5941 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 6001 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 6061 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 6121 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 6181 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 6241 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 6301 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 6361 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 6421 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 6481 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 6541 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 6601 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 6661 tccgcgcaca tttccccgaa aagtgccacc tgacgtc //