LOCUS V023060 6764 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V023060 VERSION V023060 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..2254 /label="Alg3(NM_001142363)" /note="Alg3(NM_001142363)" /gene="Alg3" CDS 2261..2326 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2364..2588 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2634..3062 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3076..3405 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3472..4266 /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 4440..4561 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4610..4626) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4634..4650 /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(4658..4688) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4703..4724 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5012..5597) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5768..6628) /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(6629..6733) /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 atggctgctg ggctgcggaa 961 acgcggcctg ccgggttctg tcgtccagcc ggcaggaatc tggaaacaat ggctacagcg 1021 ggcctggcaa gaacgctatc tactgctgcg ggagcctcgc tacaccttga tggtggcctc 1081 ctgcctctgc ctagcagagg tgggcatcac cttctgggtc attcacaggg tggcatacac 1141 agagattgac tggaaggcct acatggccca ggtggagggt ttcatcaatg gcacatatga 1201 ctacacacag ctgcagggtg acaccggacc tcttgtctac ccagctggtt ttctgtacat 1261 ctttacgggg ctttactatg ctacggaccg gggcactgac atccctacgg cccagaacat 1321 ctttgctgta ctctaccttg tcaccttggt gcttgttttc ttgatatacc accagacctc 1381 caaggtgcct cccttcgtct ttttcttcat gtgctgtgcc tcttaccgtg tccactccat 1441 ctttgtgctg cggctcttca acgacccggt agctatggca ttgctcttcc tcagcgtcaa 1501 cctcttcctg gcgcagcgct ggagctgggg ctgctgctgt ttcagcctgg ccgtctcggt 1561 gaagatgaac gtgctccttt tcgccccagg attgctattc cttcttctta ctcagttcgg 1621 cttccgtggc gccctcccca agctggccct ctgcgctgct cttcaggtgg tgctgggatt 1681 gcccttcctg ttagagaacc ccgttggcta cctgtcgcgc tcctttgacc ttggccgtca 1741 gtttctttac caatggacag tgaattggcg tctcctgcct gaagccatct tcctgcaccg 1801 tgccttccat ctggccctgc tggctgccca tcttagcttc ctcttgctat ttgccctctg 1861 taggtggcac agaacagggg aaagtattct gtcgctgctg aaggatcctt ccaaaaggaa 1921 agttccaccc caacccctca cacctaatca gattgtctcc gtcctcttca cctccaactt 1981 cattggcgtc tgcttcagcc gctccctcca ctcccagttt tatgtctggt acttccacac 2041 gctgccccac ctcctgtggg ccacgcccgc acgctggctc acacacctgc tcaggttgct 2101 ggtgctggga ctcatcgagc tctcctggaa tacataccca tccacaccct tcagctctgc 2161 tgccctacac ctgtgccacg ctgtcaccct gctgcagctc tggctgtccc ctcagttctt 2221 tcctaggagc gtccagccaa gcaggaaaac ccacctcgag gactacaaag accatgacgg 2281 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt 2341 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc 2401 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa 2461 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg 2521 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg 2581 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc 2641 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 2701 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 2761 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt 2821 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc 2881 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt 2941 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat 3001 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa 3061 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc 3121 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc 3181 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg 3241 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat 3301 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc 3361 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct 3421 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa 3481 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac 3541 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg 3601 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag 3661 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt 3721 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg 3781 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg 3841 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga 3901 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag 3961 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat 4021 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt 4081 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg 4141 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt 4201 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc 4261 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac 4321 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg 4381 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca 4441 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 4501 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 4561 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt 4621 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 4681 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 4741 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 4801 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 4861 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 4921 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 4981 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 5041 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 5101 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 5161 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 5221 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 5281 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 5341 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 5401 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 5461 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 5521 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca 5581 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 5641 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 5701 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt 5761 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt 5821 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat 5881 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag 5941 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct 6001 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt 6061 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg 6121 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca 6181 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt 6241 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat 6301 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac 6361 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa 6421 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt 6481 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt 6541 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa 6601 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt 6661 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 6721 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc //