LOCUS V039348 6334 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V039348 VERSION V039348 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 964..1824 /label="Tgif1(NM_001015020)" /note="Tgif1(NM_001015020)" /gene="Tgif1" CDS 1831..1896 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1934..2158 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2204..2632 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2646..2975 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3042..3836 /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 4010..4131 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4180..4196) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4204..4220 /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(4228..4258) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4273..4294 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4582..5167) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5338..6198) /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(6199..6303) /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 atccactagt ccagtgtggt ggaattcgcc 961 accatgattt acacgagcag gaggtgtgca ttggcggggt ccggctggcc ttcgcgtcac 1021 ggtgttgttg cagcagcatc cggcagtgac tctgaggatg aagacagtat ggacagtccc 1081 ttggaccttt catcgtcagc ggcctctggc aagagaagga ggagaggcaa tctgcccaaa 1141 gagtccgtcc agattcttcg agactggctg tatgaacaca gatacaacgc atacccctca 1201 gagcaagaga aagccctgct gtcgcagcag acacacctgt ccacactaca ggtctgtaac 1261 tggttcatca acgcccgtcg caggctcctt cctgacatgc tgagaaagga tggcaaagac 1321 ccaaatcagt tcaccatttc ccgccgaggg gccaagatct cagaagctag ctctattgaa 1381 gctgcaatgg gtatcaaaaa cttcatgcca actctagaag agagcccatt tcattcctgc 1441 gtagttggac ccaacccaac cctagggaga ccagtgtctc ccaaacctcc ctccccggga 1501 cccatcttgg ctcgcccctc agtgatctgc cataccactg tgactgcatt gaaggacggg 1561 cctttctctc tctgccaggc agtcagtgtg ggacatagta cagatgtaca gcaggtagca 1621 cccagcaact ttacagacgc ctctctcagg tacccagagg acacttgcaa atctggaccc 1681 agtccaaatc ctcagagtgg tcttttcaac actcctcccc cgactccgcc agacctcaac 1741 caggatttta gtggatttca gctcctggtg gacgttgcgc tcaagcaggc agcagagatg 1801 gagcttcagg ccaaactcac agctctcgag gactacaaag accatgacgg tgattataaa 1861 gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt ttaaacccgc 1921 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg 1981 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt 2041 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc 2101 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct 2161 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc 2221 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc 2281 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 2341 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc 2401 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg 2461 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact 2521 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt 2581 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt 2641 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 2701 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag 2761 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc 2821 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 2881 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt 2941 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca 3001 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat 3061 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac 3121 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc 3181 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc 3241 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag 3301 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc 3361 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg 3421 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc 3481 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc 3541 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga 3601 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca 3661 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg 3721 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg 3781 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg 3841 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga 3901 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg 3961 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat 4021 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 4081 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 4141 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg 4201 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 4261 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 4321 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 4381 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 4441 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 4501 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 4561 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 4621 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 4681 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 4741 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 4801 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 4861 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 4921 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 4981 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg 5041 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 5101 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 5161 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 5221 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 5281 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 5341 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 5401 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 5461 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca 5521 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 5581 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 5641 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 5701 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 5761 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 5821 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 5881 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 5941 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat 6001 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 6061 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 6121 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 6181 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 6241 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 6301 gcgcacattt ccccgaaaag tgccacctga cgtc //