LOCUS V025998 6464 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025998 VERSION V025998 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..1954 /label="HLA-G(NM_001384290)" /note="HLA-G(NM_001384290)" /gene="HLA-G" CDS 1961..2026 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2064..2288 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2334..2762 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2776..3105 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3172..3966 /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 4140..4261 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4310..4326) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4334..4350 /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(4358..4388) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4403..4424 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4712..5297) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5468..6328) /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(6329..6433) /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 atggtggtca tggcgccccg 961 aaccctcttc ctgctgctct cgggggccct gaccctgacc gagacctggg cgggctccca 1021 ctccatgagg tatttcagcg ccgccgtgtc ccggcccggc cgcggggagc cccgcttcat 1081 cgccatgggc tacgtggacg acacgcagtt cgtgcggttc gacagcgact cggcgtgtcc 1141 gaggatggag ccgcgggcgc cgtgggtgga gcaggagggg ccggagtatt gggaagagga 1201 gacacggaac accaaggccc acgcacagac tgacagaatg aacctgcaga ccctgcgcgg 1261 ctactacaac cagagcgagg ccagttctca caccctccag tggatgattg gctgcgacct 1321 ggggtccgac ggacgcctcc tccgcgggta tgaacagtat gcctacgatg gcaaggatta 1381 cctcgccctg aacgaggacc tgcgctcctg gaccgcagcg gacactgcgg ctcagatctc 1441 caagcgcaag tgtgaggcgg ccaatgtggc tgaacaaagg agagcctacc tggagggcac 1501 gtgcgtggag tggctccaca gatacctgga gaacgggaag gagatgctgc agcgcgcgga 1561 cccccccaag acacacgtga cccaccaccc tgtctttgac tatgaggcca ccctgaggtg 1621 ctgggccctg ggcttctacc ctgcggagat catactgacc tggcagcggg atggggagga 1681 ccagacccag gacgtggagc tcgtggagac caggcctgca ggggatggaa ccttccagaa 1741 gtgggcagct gtggtggtgc cttctggaga ggagcagaga tacacgtgcc atgtgcagca 1801 tgaggggctg ccggagcccc tcatgctgag atggaagcag tcttccctgc ccaccatccc 1861 catcatgggt atcgttgctg gcctggttgt ccttgcagct gtagtcactg gagctgcggt 1921 cgctgctgtg ctgtggagaa agaagagctc agatctcgag gactacaaag accatgacgg 1981 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt 2041 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc 2101 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa 2161 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg 2221 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg 2281 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc 2341 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 2401 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 2461 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt 2521 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc 2581 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt 2641 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat 2701 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa 2761 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc 2821 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc 2881 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg 2941 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat 3001 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc 3061 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct 3121 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa 3181 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac 3241 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg 3301 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag 3361 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt 3421 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg 3481 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg 3541 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga 3601 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag 3661 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat 3721 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt 3781 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg 3841 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt 3901 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc 3961 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac 4021 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg 4081 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca 4141 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 4201 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 4261 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt 4321 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 4381 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 4441 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 4501 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 4561 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 4621 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 4681 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 4741 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 4801 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 4861 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 4921 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 4981 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 5041 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 5101 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 5161 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 5221 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca 5281 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 5341 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 5401 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt 5461 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt 5521 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat 5581 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag 5641 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct 5701 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt 5761 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg 5821 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca 5881 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt 5941 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat 6001 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac 6061 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa 6121 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt 6181 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt 6241 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa 6301 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt 6361 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 6421 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc //