LOCUS V024096 7087 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024096 VERSION V024096 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..1833 /label="AQP4(NM_004028)" /note="AQP4(NM_004028)" /gene="AQP4" CDS 1849..1905 /codon_start=1 /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /product="2A peptide from porcine teschovirus-1 polyprotein" /transl_table=1 /translation="ATNFSLLKQAGDVEENPGP" /label="P2A" CDS 1906..2616 /codon_start=1 /note="mammalian codon-optimized" /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /transl_table=1 /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK*" /label="mCherry" polyA_signal 2654..2878 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2924..3352 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3366..3695 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3762..4556 /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 4730..4851 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4900..4916) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4924..4940 /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(4948..4978) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4993..5014 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5302..5887) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6058..6918) /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(6919..7023) /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 atggtggctt tcaaaggggt ctggactcaa 961 gctttctgga aagcagtcac agcggaattt ctggccatgc ttatttttgt tctcctcagc 1021 ctgggatcca ccatcaactg gggtggaaca gaaaagcctt taccggtcga catggttctc 1081 atctcccttt gctttggact cagcattgca accatggtgc agtgctttgg ccatatcagc 1141 ggtggccaca tcaaccctgc agtgactgtg gccatggtgt gcaccaggaa gatcagcatc 1201 gccaagtctg tcttctacat cgcagcccag tgcctggggg ccatcattgg agcaggaatc 1261 ctctatctgg tcacacctcc cagtgtggtg ggaggcctgg gagtcaccat ggttcatgga 1321 aatcttaccg ctggtcatgg tctcctggtt gagttgataa tcacatttca attggtgttt 1381 actatctttg ccagctgtga ttccaaacgg actgatgtca ctggctcaat agctttagca 1441 attggatttt ctgttgcaat tggacattta tttgcaatca attatactgg tgccagcatg 1501 aatcccgccc gatcctttgg acctgcagtt atcatgggaa attgggaaaa ccattggata 1561 tattgggttg ggcccatcat aggagctgtc ctcgctggtg gcctttatga gtatgtcttc 1621 tgtccagatg ttgaattcaa acgtcgtttt aaagaagcct tcagcaaagc tgcccagcaa 1681 acaaaaggaa gctacatgga ggtggaggac aacaggagtc aggtagagac ggatgacctg 1741 attctaaaac ctggagtggt gcatgtgatt gacgttgacc ggggagagga gaagaagggg 1801 aaagaccaat ctggagaggt attgtcttca gtactcgagg gaagcggagc tactaacttc 1861 agcctgctga agcaggctgg agacgtggag gagaaccctg gacctatggt gagcaagggc 1921 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc 1981 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc 2041 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc 2101 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc 2161 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag 2221 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac 2281 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc 2341 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag 2401 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc 2461 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac 2521 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc 2581 cactccaccg gcggcatgga cgagctgtac aagtaatcta gagggcccgt ttaaacccgc 2641 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg 2701 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt 2761 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc 2821 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct 2881 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc 2941 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc 3001 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 3061 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc 3121 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg 3181 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact 3241 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt 3301 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt 3361 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 3421 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag 3481 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc 3541 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 3601 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt 3661 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca 3721 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat 3781 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac 3841 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc 3901 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc 3961 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag 4021 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc 4081 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg 4141 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc 4201 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc 4261 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga 4321 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca 4381 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg 4441 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg 4501 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg 4561 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga 4621 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg 4681 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat 4741 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 4801 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 4861 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg 4921 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 4981 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 5041 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 5101 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 5161 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 5221 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 5281 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 5341 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 5401 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 5461 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 5521 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 5581 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 5641 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 5701 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg 5761 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 5821 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 5881 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 5941 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 6001 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 6061 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 6121 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 6181 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca 6241 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 6301 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 6361 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 6421 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 6481 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 6541 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 6601 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 6661 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat 6721 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 6781 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 6841 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 6901 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 6961 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 7021 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc 7081 ctatggt //