LOCUS Exported 5903 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses V5- and SBP-tagged PTB1 in mammalian cells. ACCESSION . VERSION . KEYWORDS pT7-V5-SBP-C1-HsPTB1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5903) TITLE Izaurralde lab hnRNP JOURNAL Unpublished REFERENCE 2 (bases 1 to 5903) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5903 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 613..632 /label=T7 /note="T7 promoter, forward primer" promoter 613..631 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 659..668 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 668..709 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" CDS 731..844 /codon_start=1 /product="streptavidin-binding peptide" /label=SBP /note="selected from a peptide library; binds streptavidin with nanomolar affinity (Keefe et al., 2001)" /translation="MDEKTTGWGGGHVVEGLAGELEQLRARLEHHPQGQREP" polyA_signal 2691..2812 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2728..2747) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2782..2801 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2819..3274) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2956..2977) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3168..3187 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3301..3405 /gene="bla" /label=AmpR promoter promoter 3407..3764 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3430..3450) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3615..3750 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3677..3696 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3799..4593 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3853..3872) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4463..4482 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4781..4800) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4825..4872 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5201..5789 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5690..5709 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg 601 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc 661 caccatgggt aagcctatcc ctaaccctct cctcggtctc gattctacgc ccggaagcgg 721 tagcggtacg atggacgaga agaccaccgg ctgggggggc ggccacgtgg tggagggcct 781 ggccggcgag ctggagcagc tgcgggccag gctggagcac caccctcagg gccagcggga 841 gcccagatct cgagacatgg acggcattgt cccagatata gccgttggta caaagcgggg 901 atctgacgag cttttctcta cttgtgtcac taacggaccg tttatcatga gcagcaactc 961 ggcttctgca gcaaacggaa atgacagcaa gaagttcaaa ggtgacagcc gaagtgcagg 1021 cgtcccctct agagtgatcc acatccggaa gctccccatc gacgtcacgg agggggaagt 1081 catctccctg gggctgccct ttgggaaggt caccaacctc ctgatgctga aggggaaaaa 1141 ccaggccttc atcgagatga acacggagga ggctgccaac accatggtga actactacac 1201 ctcggtgacc cctgtgctgc gcggccagcc catctacatc cagttctcta accacaagga 1261 gctgaagacc gacagctctc ccaaccaggc gcgggcccag gcggccctgc aggcggtgaa 1321 ctcggtccag tcggggaacc tggccttggc tgcctcggcg gcggccgtgg acgcagggat 1381 ggcgatggcc gggcagagcc ctgtgctcag gatcatcgtg gagaacctct tctaccctgt 1441 gaccctggat gtgctgcacc agattttctc caagttcggc acagtgttga agatcatcac 1501 cttcaccaag aacaaccagt tccaggccct gctgcagtat gcggaccccg tgagcgccca 1561 gcacgccaag ctgtcgctgg acgggcagaa catctacaac gcctgctgca cgctgcgcat 1621 cgacttttcc aagctcacca gcctcaacgt caagtacaac aatgacaaga gccgtgacta 1681 cacacgccca gacctgcctt ccggggacag ccagccctcg ctggaccaga ccatggccgc 1741 ggccttcggt gcacctggta taatctcagc ctctccgtat gcaggagctg gtttccctcc 1801 cacctttgcc attcctcaag ctgcaggcct ttccgttccg aacgtccacg gcgccctggc 1861 ccccctggcc atcccctcgg cggcggcggc agctgcggcg gcaggtcgga tcgccatccc 1921 gggcctggcg ggggcaggaa attctgtatt gctggtcagc aacctcaacc cagagagagt 1981 cacaccccaa agcctcttta ttcttttcgg cgtctacggt gacgtgcagc gcgtgaagat 2041 cctgttcaat aagaaggaga acgccctagt gcagatggcg gacggcaacc aggcccagct 2101 ggccatgagc cacctgaacg ggcacaagct gcacgggaag ccgatccgca tcacgctctc 2161 gaagcaccag aacgtgcagc tgccccgcga gggccaggag gaccagggcc tgaccaagga 2221 ctacggcaac tcacccctgc accgcttcaa gaagccgggc tccaagaact tccagaacat 2281 attcccgccc tcggccactc tgcacctctc caacatcccg ccctcagtct ccgaggagga 2341 tctcaaggtc ctgttttcca gcaatggggg cgtcgtcaaa ggattcaagt tcttccagaa 2401 ggaccgcaag atggcactga tccagatggg ctccgtggag gaggcggtcc aggccctcat 2461 tgacctgcac aaccacgacc tcggggagaa ccaccacctg cgggtctcct tctccaagtc 2521 caccatctag gaattctgca gtcgacggta ccgcgggccc gggatccacc ggatctagat 2581 aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc 2641 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta 2701 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 2761 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa 2821 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt 2881 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata 2941 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac 3001 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa 3061 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc 3121 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg 3181 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca 3241 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg 3301 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga 3361 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga 3421 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca 3481 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct 3541 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc 3601 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg 3661 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc 3721 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca 3781 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct 3841 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc 3901 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc 3961 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc 4021 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg 4081 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc 4141 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac 4201 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat 4261 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc 4321 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg 4381 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg 4441 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc 4501 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc 4561 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg 4621 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa 4681 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc 4741 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa 4801 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg 4861 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac 4921 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa 4981 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct 5041 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct 5101 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc 5161 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc 5221 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg 5281 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa 5341 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc 5401 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt 5461 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa 5521 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc 5581 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc 5641 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct 5701 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat 5761 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc 5821 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg 5881 ataaccgtat taccgccatg cat //