LOCUS V042128 6094 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V042128 VERSION V042128 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..1584 /label="Rab8a(NM_023126)" /note="Rab8a(NM_023126)" /gene="Rab8a" CDS 1591..1656 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1694..1918 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1964..2392 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2406..2735 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2802..3596 /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 3770..3891 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3940..3956) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3964..3980 /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(3988..4018) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4033..4054 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4342..4927) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5098..5958) /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(5959..6063) /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 accatggcga agacctacga ttacctgttc aagctgctgc tgatcgggga ctcgggggta 1021 gggaagacct gtgtcctgtt ccgcttctcc gaggacgcct tcaactccac attcatctct 1081 accataggaa ttgactttaa aattaggacc atagagctcg atggcaagag gattaaactg 1141 cagatatggg acacggccgg ccaggagcgg tttcgaacaa tcacgacagc ctactacagg 1201 ggtgccatgg gtatcatgct ggtctacgac attaccaatg agaagtcctt tgacaacatc 1261 cggaattgga ttcggaacat tgaagagcat gcctctgcag acgtggagaa gatgatactg 1321 gggaataagt gtgatgtgaa tgacaagaga caggtgtcca aggaacgggg agaaaagctg 1381 gcactcgact atgggatcaa gttcatggag accagtgcaa aggccaacat caatgtggag 1441 aatgcatttt tcactcttgc cagggatatc aaagcaaaaa tggacaaaaa attggaaggg 1501 aacagcccgc aggggagcag ccatggagtc aagatcacag tggagcagca gaagaggacc 1561 agcttcttcc ggtgcagtct cctgctcgag gactacaaag accatgacgg tgattataaa 1621 gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt ttaaacccgc 1681 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg 1741 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt 1801 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc 1861 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct 1921 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc 1981 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc 2041 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 2101 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc 2161 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg 2221 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact 2281 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt 2341 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt 2401 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 2461 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag 2521 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc 2581 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 2641 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt 2701 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca 2761 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat 2821 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac 2881 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc 2941 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc 3001 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag 3061 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc 3121 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg 3181 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc 3241 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc 3301 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga 3361 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca 3421 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg 3481 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg 3541 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg 3601 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga 3661 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg 3721 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat 3781 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 3841 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 3901 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg 3961 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 4021 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 4081 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 4141 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 4201 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 4261 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 4321 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 4381 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 4441 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 4501 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 4561 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 4621 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 4681 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 4741 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg 4801 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 4861 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 4921 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 4981 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 5041 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 5101 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 5161 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 5221 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca 5281 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 5341 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 5401 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 5461 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 5521 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 5581 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 5641 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 5701 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat 5761 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 5821 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 5881 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 5941 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 6001 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 6061 gcgcacattt ccccgaaaag tgccacctga cgtc //