LOCUS V025824 7034 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025824 VERSION V025824 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..2524 /label="ALPI(NM_001631)" /note="ALPI(NM_001631)" /gene="ALPI" CDS 2531..2596 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2634..2858 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2904..3332 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3346..3675 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3742..4536 /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 4710..4831 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4880..4896) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4904..4920 /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(4928..4958) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4973..4994 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5282..5867) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6038..6898) /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(6899..7003) /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 atgcaggggc cctgggtgct 961 gctgctgctg ggcctgaggc tacagctctc cctgggcgtc atcccagctg aggaggagaa 1021 cccggccttc tggaaccgcc aggcagctga ggccctggat gctgccaaga agctgcagcc 1081 catccagaag gtcgccaaga acctcatcct cttcctgggc gatgggttgg gggtgcccac 1141 ggtgacagcc accaggatcc taaaggggca gaagaatggc aaactggggc ctgagacgcc 1201 cctggccatg gaccgcttcc catacctggc tctgtccaag acatacaatg tggacagaca 1261 ggtgccagac agcgcagcca cagccacggc ctacctgtgc ggggtcaagg ccaacttcca 1321 gaccatcggc ttgagtgcag ccgcccgctt taaccagtgc aacacgacac gcggcaatga 1381 ggtcatctcc gtgatgaacc gggccaagca agcaggaaag tcagtaggag tggtgaccac 1441 cacacgggtg cagcacgcct cgccagccgg cacctacgca cacacagtga accgcaactg 1501 gtactcagat gctgacatgc ctgcctcagc ccgccaggag gggtgccagg acatcgccac 1561 tcagctcatc tccaacatgg acattgacgt gatccttggc ggaggccgca agtacatgtt 1621 tcccatgggg accccagacc ctgagtaccc agctgatgcc agccagaatg gaatcaggct 1681 ggacgggaag aacctggtgc aggaatggct ggcaaagcac cagggtgcct ggtatgtgtg 1741 gaaccgcact gagctcatgc aggcgtccct ggaccagtct gtgacccatc tcatgggcct 1801 ctttgagccc ggagacacga aatatgagat ccaccgagac cccacactgg acccctccct 1861 gatggagatg acagaggctg ccctgcgcct gctgagcagg aacccccgcg gcttctacct 1921 ctttgtggag ggcggccgca tcgaccatgg tcatcatgag ggtgtggctt accaggcact 1981 cactgaggcg gtcatgttcg acgacgccat tgagagggcg ggccagctca ccagcgagga 2041 ggacacgctg accctcgtca ccgctgacca ctcccatgtc ttctcctttg gtggctacac 2101 cttgcgaggg agctccatct tcgggttggc ccccagcaag gctcaggaca gcaaagccta 2161 cacgtccatc ctgtacggca atggcccggg ctacgtgttc aactcaggcg tgcgaccaga 2221 cgtgaatgag agcgagagcg ggagccccga ttaccagcag caggcggcgg tgcccctgtc 2281 gtccgagacc cacggaggcg aagacgtggc ggtgtttgcg cgcggcccgc aggcgcacct 2341 ggtgcatggt gtgcaggagc agagcttcgt agcgcatgtc atggccttcg ctgcctgtct 2401 ggagccctac acggcctgcg acctggcgcc tcccgcctgc accaccgacg ccgcgcaccc 2461 agttgccgcg tcgctgccac tgctggccgg gaccctgctg ctgctggggg cgtccgctgc 2521 tcccctcgag gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa 2581 ggatgacgat gacaagtagt gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc 2641 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg 2701 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag 2761 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga 2821 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag 2881 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt 2941 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc 3001 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg 3061 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta 3121 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt 3181 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat 3241 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa 3301 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg 3361 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 3421 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 3481 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact 3541 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 3601 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 3661 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag 3721 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc 3781 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat 3841 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg 3901 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg 3961 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta 4021 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta 4081 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc 4141 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc 4201 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg 4261 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg 4321 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt 4381 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc 4441 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc 4501 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga 4561 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg 4621 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg 4681 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca 4741 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt 4801 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct 4861 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 4921 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 4981 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 5041 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 5101 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 5161 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 5221 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 5281 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 5341 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 5401 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 5461 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 5521 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 5581 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 5641 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 5701 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 5761 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt 5821 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 5881 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 5941 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 6001 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 6061 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 6121 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 6181 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 6241 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 6301 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 6361 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 6421 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 6481 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 6541 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 6601 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 6661 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 6721 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 6781 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 6841 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 6901 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 6961 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 7021 tgccacctga cgtc //