LOCUS V022665 6434 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022665 VERSION V022665 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..1924 /label="GNRHR(NM_000406)" /note="GNRHR(NM_000406)" /gene="GNRHR" CDS 1931..1996 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2034..2258 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2304..2732 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2746..3075 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3142..3936 /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 4110..4231 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4280..4296) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4304..4320 /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(4328..4358) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4373..4394 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4682..5267) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5438..6298) /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(6299..6403) /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 atggcaaaca gtgcctctcc 961 tgaacagaat caaaatcact gttcagccat caacaacagc atcccactga tgcagggcaa 1021 cctccccact ctgaccttgt ctggaaagat ccgagtgacg gttactttct tcctttttct 1081 gctctctgcg acctttaatg cttctttctt gttgaaactt cagaagtgga cacagaagaa 1141 agagaaaggg aaaaagctct caagaatgaa gctgctctta aaacatctga ccttagccaa 1201 cctgttggag actctgattg tcatgccact ggatgggatg tggaacatta cagtccaatg 1261 gtatgctgga gagttactct gcaaagttct cagttatcta aagcttttct ccatgtatgc 1321 cccagccttc atgatggtgg tgatcagcct ggaccgctcc ctggctatca cgaggcccct 1381 agctttgaaa agcaacagca aagtcggaca gtccatggtt ggcctggcct ggatcctcag 1441 tagtgtcttt gcaggaccac agttatacat cttcaggatg attcatctag cagacagctc 1501 tggacagaca aaagttttct ctcaatgtgt aacacactgc agtttttcac aatggtggca 1561 tcaagcattt tataactttt tcaccttcag ctgcctcttc atcatccctc ttttcatcat 1621 gctgatctgc aatgcaaaaa tcatcttcac cctgacacgg gtccttcatc aggaccccca 1681 cgaactacaa ctgaatcagt ccaagaacaa tataccaaga gcacggctga agactctaaa 1741 aatgacggtt gcatttgcca cttcatttac tgtctgctgg actccctact atgtcctagg 1801 aatttggtat tggtttgatc ctgaaatgtt aaacaggttg tcagacccag taaatcactt 1861 cttctttctc tttgcctttt taaacccatg ctttgatcca cttatctatg gatatttttc 1921 tctgctcgag gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa 1981 ggatgacgat gacaagtagt gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc 2041 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg 2101 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag 2161 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga 2221 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag 2281 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt 2341 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc 2401 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg 2461 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta 2521 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt 2581 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat 2641 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa 2701 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg 2761 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 2821 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 2881 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact 2941 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 3001 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 3061 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag 3121 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc 3181 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat 3241 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg 3301 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg 3361 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta 3421 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta 3481 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc 3541 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc 3601 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg 3661 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg 3721 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt 3781 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc 3841 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc 3901 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga 3961 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg 4021 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg 4081 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca 4141 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt 4201 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct 4261 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 4321 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 4381 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 4441 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 4501 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 4561 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 4621 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 4681 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 4741 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 4801 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 4861 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 4921 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 4981 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 5041 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 5101 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 5161 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt 5221 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 5281 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 5341 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 5401 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 5461 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 5521 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 5581 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 5641 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 5701 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 5761 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 5821 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 5881 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 5941 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 6001 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 6061 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 6121 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 6181 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 6241 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 6301 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 6361 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 6421 tgccacctga cgtc //