LOCUS V025486 7115 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025486 VERSION V025486 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..2605 /label="Ptbp1(NM_001077363)" /note="Ptbp1(NM_001077363)" /gene="Ptbp1" CDS 2612..2677 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2715..2939 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2985..3413 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3427..3756 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3823..4617 /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 4791..4912 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4961..4977) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4985..5001 /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(5009..5039) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5054..5075 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5363..5948) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6119..6979) /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(6980..7084) /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 atggacggca tcgtcccaga 961 catagcagtc ggtacaaagc ggggatccga cgagctcttc tccacgtgtg tcagcaacgg 1021 ccccttcatc atgagcagct ctgcctcagc agccaatgga aacgatagca agaagttcaa 1081 aggtgacaac aggagcgcag gagtcccttc cagagtcatc catgtcagaa agctgcccag 1141 cgatgtcact gagggcgagg tcatctccct agggctgccc tttggaaagg ttaccaacct 1201 tctcatgctg aaggggaaga accaggcctt cattgagatg aacacagagg aggctgccaa 1261 cactatggtt aactactata catcggtggc gccagtgctt cgtggacagc ccatctacat 1321 ccagttctcc aaccacaaag agctcaagac cgacagctcg cccaaccagg cacgtgccca 1381 ggcagccctg caggctgtaa actccgtcca gtctggaaac ctggccttgg cagcgtccgc 1441 tgctgccgtg gatgcaggaa tggcaatggc agggcagagc ccagtgctca ggatcattgt 1501 ggaaaacctt ttctacccag tgaccctgga cgtgctgcac cagatcttct ctaagtttgg 1561 caccgtcctg aagatcatca cgttcaccaa gaacaaccag ttccaggcgc tgctgcagta 1621 tgctgaccct gtgagcgccc agcatgccaa gctgtccctg gatggccaga acatctacaa 1681 cgcctgctgc acgctgcgca tcgacttctc caagctcacc agtctcaatg tcaagtacaa 1741 caatgataag agcagagact acactcgacc tgacctgccc tctggagaca gccagccttc 1801 actagaccag accatggcag cagcctttgg tgcgcccggc ataatgtcag cctctccgta 1861 tgcaggagcc gggttccctc ccacctttgc catccctcag gccgcaggcc tctctgtccc 1921 taatgtccat ggagccttgg cccccctggc catcccgtct gctgctgctg ctgctgcggc 1981 cagccgcatt gccatcccag ggttggcagg tgctgggaat tctgtccttt tggtcagcaa 2041 tctgaaccct gagagagtca caccccaaag cctctttatt ctcttcggcg tctacggtga 2101 tgtgcagcgg gtgaagatcc tgttcaataa gaaggagaac gcacttgtgc agatggcaga 2161 cggcagccag gcccagctgg ccatgagcca cctgaacggg cacaagctgc acgggaagtc 2221 agtgcgcatt acactgtcca agcatcagag tgtgcagctg cctcgggagg gtcaggagga 2281 ccagggcctg accaaggact atggcagctc cccgctgcac cgcttcaaga aaccaggctc 2341 caagaacttc cagaacatct ttccaccctc agctaccctg cacctctcca acatcccgcc 2401 ctctgtgtca gaggacgacc tcaagagcct cttctccagc aacggtggtg tggtcaaagg 2461 cttcaagttc ttccagaagg accgcaagat ggcactgatc cagatgggct ctgtggagga 2521 ggctgtgcag gcgctgattg aactgcacaa ccatgacctg ggcgagaacc accacctgcg 2581 agtgtccttt tccaagtcca ccatcctcga ggactacaaa gaccatgacg gtgattataa 2641 agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg tttaaacccg 2701 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt 2761 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat 2821 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag 2881 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc 2941 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg 3001 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc 3061 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc 3121 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct 3181 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac 3241 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac 3301 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat 3361 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg 3421 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg 3481 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca 3541 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact 3601 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta 3661 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag 3721 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc 3781 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga 3841 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa 3901 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt 3961 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg 4021 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa 4081 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac 4141 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt 4201 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact 4261 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg 4321 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg 4381 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc 4441 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt 4501 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc 4561 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg 4621 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg 4681 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct 4741 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta 4801 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 4861 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct 4921 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt 4981 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag 5041 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt 5101 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag 5161 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 5221 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 5281 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 5341 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 5401 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 5461 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 5521 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 5581 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 5641 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 5701 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 5761 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 5821 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 5881 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 5941 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 6001 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 6061 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 6121 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 6181 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 6241 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 6301 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 6361 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 6421 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 6481 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 6541 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 6601 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 6661 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 6721 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 6781 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 6841 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 6901 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 6961 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 7021 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 7081 cgcgcacatt tccccgaaaa gtgccacctg acgtc //