LOCUS V040627 6721 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040627 VERSION V040627 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..2211 /label="ALG2(NM_033087)" /note="ALG2(NM_033087)" /gene="ALG2" CDS 2218..2283 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2321..2545 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2591..3019 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3033..3362 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3429..4223 /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 4397..4518 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4567..4583) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4591..4607 /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(4615..4645) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4660..4681 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4969..5554) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5725..6585) /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(6586..6690) /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 accatggcgg aggagcaggg ccgggaacgg gactcggttc ccaagccgtc ggtgctgttc 1021 ctccacccag acctgggcgt gggcggcgct gagcggctgg tgttggacgc ggcgctggcg 1081 ctgcaggcgc gcgggtgtag cgtgaagatc tggacagcgc actacgaccc gggccactgt 1141 ttcgccgaga gccgcgagct accggtgcgc tgtgccgggg actggctgcc gcgaggcctg 1201 ggctggggcg gccgcggcgc cgccgtctgc gcctacgtgc gcatggtttt cctggcgctc 1261 tacgtgctgt tcctcgccga cgaggagttc gacgtggtag tgtgcgacca ggtgtctgcc 1321 tgtatcccag tgttcaggct ggctagacgg cggaagaaga tcctatttta ctgtcacttc 1381 ccagatctgc ttctcaccaa gagagattct tttcttaaac gactatacag ggccccaatt 1441 gactggatag aggaatacac cacaggcatg gcagactgca tcttagtcaa cagccagttc 1501 acagctgctg tttttaagga aacattcaag tccctgtctc acatagaccc tgatgtcctc 1561 tatccatctc taaatgtcac cagctttgac tcagttgttc ctgaaaagct ggatgaccta 1621 gtccccaagg ggaaaaaatt cctgctgctc tccatcaaca gatacgaaag gaagaaaaat 1681 ctgactttgg cactggaagc cctagtacag ctgcgtggaa gattgacatc ccaagattgg 1741 gagagggttc atctgatcgt ggcaggtggt tatgacgaga gagtcctgga gaatgtggaa 1801 cattatcagg aattgaagaa aatggtccaa cagtccgacc ttggccagta tgtgaccttc 1861 ttgaggtctt tctcagacaa acagaaaatc tccctcctcc acagctgcac gtgtgtgctt 1921 tacacaccaa gcaatgagca ctttggcatt gtccctctgg aagccatgta catgcagtgc 1981 ccagtcattg ctgttaattc gggtggaccc ttggagtcca ttgaccacag tgtcacaggg 2041 tttctgtgtg agcctgaccc ggtgcacttc tcagaagcaa tagaaaagtt catccgtgaa 2101 ccttccttaa aagccaccat gggcctggct ggaagagcca gagtgaagga aaaattttcc 2161 cctgaagcat ttacagaaca gctctaccga tatgttacca aactgctggt actcgaggac 2221 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac 2281 aagtagtgag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag 2341 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact 2401 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt 2461 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat 2521 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg 2581 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc 2641 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc 2701 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg 2761 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca 2821 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc 2881 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct 2941 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa 3001 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc 3061 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt 3121 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 3181 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg 3241 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct 3301 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca 3361 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat 3421 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga 3481 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc 3541 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga 3601 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg 3661 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc 3721 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg 3781 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga 3841 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc 3901 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca 3961 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg 4021 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct 4081 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg 4141 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc 4201 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac 4261 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt 4321 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga 4381 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag 4441 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 4501 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta 4561 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat 4621 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt 4681 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta 4741 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc 4801 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 4861 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 4921 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 4981 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 5041 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 5101 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 5161 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 5221 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 5281 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 5341 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 5401 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 5461 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt ttgtttgcaa 5521 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 5581 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 5641 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 5701 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 5761 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 5821 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 5881 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 5941 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 6001 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 6061 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 6121 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 6181 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 6241 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 6301 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 6361 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 6421 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 6481 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 6541 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 6601 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 6661 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 6721 c //