LOCUS V023989 7367 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V023989 VERSION V023989 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 941..2857 /label="ILDR2(NM_199351)" /note="ILDR2(NM_199351)" /gene="ILDR2" CDS 2864..2929 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2967..3191 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3237..3665 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3679..4008 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4075..4869 /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 5043..5164 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5213..5229) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5237..5253 /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(5261..5291) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5306..5327 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5615..6200) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6371..7231) /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(7232..7336) /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 aagcttggta ccgagctcgg atccgccacc atggataggg tcttgctgag 961 gtggatttct ctcttctggc taacagccat ggtcgaaggc cttcaggtca cagtgcccga 1021 caagaagaag gtggccatgc tcttccagcc cactgtgctt cgctgccact tctcaacatc 1081 ctcccatcag cctgcagttg tgcagtggaa gttcaagtcc tactgccagg atcgcatggg 1141 agaatccttg ggcatgtcct ctacccgggc ccaatctctc agcaagagaa acctggaatg 1201 ggacccctac ttggattgtt tggacagcag gaggactgtt cgagtagtag cttcaaaaca 1261 gggctcgact gtcaccctgg gagatttcta caggggcaga gagatcacga ttgttcatga 1321 tgcagatctt caaattggaa agcttatgtg gggagacagc ggactctatt actgtattat 1381 caccacccca gatgacctgg aggggaaaaa tgaggactca gtggaactgc tggtgttggg 1441 caggacaggg ctgcttgctg atctcttgcc cagttttgct gtggagatta tgccagagtg 1501 ggtgtttgtt ggcctggtgc tcctgggcgt cttcctcttc ttcgtcctgg tggggatctg 1561 ctggtgccag tgctgccctc acagctgctg ctgctatgtc cgctgcccat gctgcccaga 1621 ttcctgctgc tgccctcaag ccttgtatga agcagggaaa gcagcaaagg ccgggtaccc 1681 tccctctgtc tccggtgtcc ccggccctta ctccatcccc tctgtccctt tgggaggagc 1741 cccctcatct ggcatgctga tggacaagcc gcatccacct cccttggcac caagtgactc 1801 cactggagga agccacagtg ttcgcaaagg ttaccggatc caggctgaca aagagagaga 1861 ctccatgaag gtcctgtact atgttgagaa ggagctggct cagtttgatc cagccagaag 1921 gatgagaggc agatataaca acaccatctc agaactcagc tccctacatg aggaggacag 1981 caatttccgc cagtctttcc atcagatgag aagcaagcag ttccctgtgt ctggggactt 2041 ggagagcaat cctgactatt ggtcaggtgt catgggaggc agcagtgggg caagccgcgg 2101 gccctcagcc atggagtata acaaagagga tcgagagagc ttcaggcaca gccagccgcg 2161 ctccaagtcg gagatgctgt cgcggaagaa cttcgccacg ggggtgccgg ccgtttccat 2221 ggacgagctg gcggccttcg ctgactccta cggccagcgg ccccgccggg cagacggcaa 2281 cagtcacgag gcgcggggcg ggagccgctt cgagcgctcg gagtcgcggg cgcacagcgg 2341 cttctaccag gacgactcct tggaggagta ctacggtcag cgcagccgca gccgcgagcc 2401 cctgaccgat gctgaccgcg gctgggcctt cagccccgcg cgccgcagac ccgccgagga 2461 cgcgcacctg ccgcggctgg tgagccgcac gccaggcacc gcacccaaat acgaccactc 2521 gtacctgggc agcgcgcggg agcgccaggc gcggcccgag ggcgccagcc gcggtggcag 2581 cctggagacg ccatccaagc ggagcgcgca gctcggcccg cgcagcgcct cctactacgc 2641 ttggtcgccg cccggcacct acaaggccgg ctcgtcgcag gacgaccagg aggacgcgtc 2701 cgacgacgcg ctgccgccct acagcgagct ggagctgacc cgcggcccgt cctaccgcgg 2761 ccgcgacctg ccctaccaca gcaactcgga gaagaagagg aaaaaggagc ccgccaagaa 2821 aaccaatgac tttccaacca ggatgtccct tgtggtcctc gaggactaca aagaccatga 2881 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc 2941 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg 3001 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata 3061 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt 3121 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt 3181 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc 3241 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac 3301 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt 3361 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc 3421 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc 3481 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact 3541 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg 3601 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc 3661 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca 3721 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca 3781 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc 3841 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc 3901 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta 3961 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga 4021 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt 4081 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat 4141 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag 4201 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac 4261 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac 4321 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc 4381 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg 4441 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag 4501 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat 4561 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag 4621 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc 4681 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg 4741 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg 4801 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag 4861 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat 4921 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc 4981 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc 5041 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca 5101 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat 5161 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc 5221 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 5281 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 5341 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 5401 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 5461 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 5521 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 5581 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 5641 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 5701 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 5761 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 5821 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 5881 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 5941 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 6001 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 6061 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 6121 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc 6181 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 6241 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 6301 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 6361 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 6421 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 6481 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 6541 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 6601 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 6661 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 6721 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 6781 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 6841 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 6901 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 6961 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 7021 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 7081 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 7141 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 7201 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 7261 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 7321 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc //