LOCUS V042749 7232 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V042749 VERSION V042749 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 908..1618 /codon_start=1 /note="mammalian codon-optimized" /product="enhanced monomeric blue fluorescent protein (Subach et al., 2011)" /transl_table=1 /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV ARYCDLPSKLGHKLN" /label="mTagBFP2" CDS 1649..2356 /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" CDS 2378..2752 /codon_start=1 /gene="MAP1LC3B" /note="Atg8 family member found on autophagosomes" /product="human LC3 protein" /transl_table=1 /translation="MPSEKTFKQRRTFEQRVEDVRLIREQHPTKIPVIIERYKGEKQLP VLDKTKFLVPDHVNMSELIKIIRRRLQLNANQAFFLLVNGHSMVSVSTPISEVYESEKD EDGFLYMVYASQETFGMKLSV" /label="LC3B" polyA_signal 2799..3023 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3069..3497 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3511..3840 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3907..4701 /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 4875..4996 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5045..5061) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5069..5085 /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(5093..5123) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5138..5159 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5447..6032) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6203..7063) /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(7064..7168) /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 cgccaccatg gtgtctaagg gcgaagagct gattaaggag aacatgcaca tgaagctgta 961 catggagggc accgtggaca accatcactt caagtgcaca tccgagggcg aaggcaagcc 1021 ctacgagggc acccagacca tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc 1081 cttcgacatc ctggctacta gcttcctcta cggcagcaag accttcatca accacaccca 1141 gggcatcccc gacttcttca agcagtcctt ccctgagggc ttcacatggg agagagtcac 1201 cacatacgaa gacgggggcg tgctgaccgc tacccaggac accagcctcc aggacggctg 1261 cctcatctac aacgtcaaga tcagaggggt gaacttcaca tccaacggcc ctgtgatgca 1321 gaagaaaaca ctcggctggg aggccttcac cgagacgctg taccccgctg acggcggcct 1381 ggaaggcaga aacgacatgg ccctgaagct cgtgggcggg agccatctga tcgcaaacgc 1441 caagaccaca tatagatcca agaaacccgc taagaacctc aagatgcctg gcgtctacta 1501 tgtggactac agactggaaa gaatcaagga ggccaacaac gagacctacg tcgagcagca 1561 cgaggtggca gtggccagat actgcgacct ccctagcaaa ctggggcaca agcttaatgg 1621 tggaggcgga tcaggaggcg gtggatctat ggtgagcaag ggcgaggagg ataacatggc 1681 catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga 1741 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct 1801 gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat 1861 gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc 1921 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac 1981 cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg 2041 caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc 2101 ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa 2161 gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc 2221 cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga 2281 ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat 2341 ggacgagctg tacaagtccg gactcagatc tcgagctatg ccgtcggaga agaccttcaa 2401 gcagcgccgc accttcgaac aaagagtaga agatgtccga cttattcgag agcagcatcc 2461 aaccaaaatc ccggtgataa tagaacgata caagggtgag aagcagcttc ctgttctgga 2521 taaaacaaag ttccttgtac ctgaccatgt caacatgagt gagctcatca agataattag 2581 aaggcgctta cagctcaatg ctaatcaggc cttcttcctg ttggtgaacg gacacagcat 2641 ggtcagcgtc tccacaccaa tctcagaggt gtatgagagt gagaaagatg aagatggatt 2701 cctgtacatg gtctatgcct cccaggagac gttcgggatg aaattgtcag tgtaactcga 2761 gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc 2821 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt 2881 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct 2941 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc 3001 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg 3061 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag 3121 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt 3181 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt 3241 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg 3301 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt 3361 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt 3421 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca 3481 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca 3541 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt 3601 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 3661 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 3721 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct 3781 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa 3841 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg 3901 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg 3961 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg 4021 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat 4081 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca 4141 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg 4201 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat 4261 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa 4321 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg 4381 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg 4441 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg 4501 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat 4561 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac 4621 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc 4681 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc 4741 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg 4801 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt 4861 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca 4921 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac 4981 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat 5041 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac 5101 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa 5161 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat 5221 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc 5281 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg 5341 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag 5401 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc 5461 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag 5521 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga 5581 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc 5641 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg 5701 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt 5761 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca 5821 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca 5881 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag 5941 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc 6001 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 6061 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 6121 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 6181 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga 6241 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac 6301 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg 6361 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg 6421 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt 6481 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct 6541 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 6601 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta 6661 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca 6721 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat 6781 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac 6841 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa 6901 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt 6961 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg 7021 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat 7081 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt 7141 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg 7201 acggatcggg agatctcccg atcccctatg gt //