LOCUS V045014 6304 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045014 VERSION V045014 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 202..581 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 582..785 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 830..848 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 931..1743 /label="SPI1(NM_001080547)" /note="SPI1(NM_001080547)" /gene="SPI1" CDS 1750..1776 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 1780..1806 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 1810..1836 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" polyA_signal 1871..2095 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2141..2569 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2583..2912 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2979..3773 /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 3947..4068 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4117..4133) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4141..4157 /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(4165..4195) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4210..4231 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4519..5104) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5275..6135) /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(6136..6240) /gene="bla" /label="AmpR promoter" ORIGIN 1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt 61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct 121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg 181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta 241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc 841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc 901 cactagtcca gtgtggtgga attcgccacc atgttacagg cgtgcaaaat ggaagggttt 961 cccctcgtcc cccctcagcc atcagaagac ctggtgccct atgacacgga tctataccaa 1021 cgccaaacgc acgagtatta cccctatctc agcagtgatg gggagagcca tagcgaccat 1081 tactgggact tccaccccca ccacgtgcac agcgagttcg agagcttcgc cgagaacaac 1141 ttcacggagc tccagagcgt gcagcccccg cagctgcagc agctctaccg ccacatggag 1201 ctggagcaga tgcacgtcct cgataccccc atggtgccac cccatcccag tcttggccac 1261 caggtctcct acctgccccg gatgtgcctc cagtacccat ccctgtcccc agcccagccc 1321 agctcagatg aggaggaggg cgagcggcag agccccccac tggaggtgtc tgacggcgag 1381 gcggatggcc tggagcccgg gcctgggctc ctgcctgggg agacaggcag caagaagaag 1441 atccgcctgt accagttcct gttggacctg ctccgcagcg gcgacatgaa ggacagcatc 1501 tggtgggtgg acaaggacaa gggcaccttc cagttctcgt ccaagcacaa ggaggcgctg 1561 gcgcaccgct ggggcatcca gaagggcaac cgcaagaaga tgacctacca gaagatggcg 1621 cgcgcgctgc gcaactacgg caagacgggc gaggtcaaga aggtgaagaa gaagctcacc 1681 taccagttca gcggcgaagt gctgggccgc gggggcctgg ccgagcggcg ccacccgccc 1741 cacctcgagt acccatacga cgtacctgat tatgcaactt atccttatga tgtacctgat 1801 tatgctagct atccttatga tgtcccagac tacgcatgag ggcccgttta aacccgctga 1861 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct 1921 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca 1981 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag 2041 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct 2101 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca 2161 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 2221 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 2281 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 2341 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt 2401 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 2461 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg 2521 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga 2581 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 2641 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca 2701 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc 2761 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt 2821 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag 2881 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt 2941 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc 3001 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga 3061 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt 3121 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat 3181 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg 3241 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg 3301 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc 3361 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga 3421 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag 3481 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc 3541 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg 3601 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata 3661 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg 3721 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac 3781 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc 3841 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat 3901 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc 3961 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt 4021 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat 4081 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa 4141 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg 4201 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca 4261 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg 4321 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 4381 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 4441 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 4501 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 4561 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 4621 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 4681 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 4741 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 4801 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 4861 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 4921 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 4981 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 5041 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc 5101 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg 5161 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta 5221 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca 5281 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc 5341 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc 5401 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc 5461 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat 5521 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt 5581 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc 5641 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag 5701 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt 5761 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac 5821 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg 5881 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat 5941 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc 6001 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc 6061 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa 6121 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg 6181 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg 6241 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta 6301 tggt //