LOCUS Exported 9537 bp ds-DNA circular SYN 11-OCT-2022 DEFINITION Imaging of synaptopodin in live cells. ACCESSION . VERSION . KEYWORDS Venus/Cerulean-Synaptopodin-A in pcDNA6 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9537) AUTHORS Morris T, Sue E, Geniesse C, Brieher WM, Tang VW TITLE Synaptopodin stress fiber and contractomere at the epithelial junction. JOURNAL J Cell Biol. 2022 May 2;221(5). pii: 213132. doi: 10.1083/jcb.202011162. Epub 2022 Apr 13. PUBMED 35416930 REFERENCE 2 (bases 1 to 9537) AUTHORS . TITLE Direct Submission JOURNAL Exported Oct 11, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9537 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 914..1633 /codon_start=1 /product="Venus YFP with monomerizing A206K mutation (Nagai et al., 2002; Kremers et al., 2006)" /label=mVenus /note="mammalian codon-optimized" /translation="MSVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLT LKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKD DGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGI KANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVL LEFVTAAGITLGMDELYK" CDS 1961..2677 /codon_start=1 /product="enhanced monomeric variant of CFP (Rizzo et al., 2004)" /label=mCerulean /note=" mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 5408..5437 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 5453..5470 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(5493..5510) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5499..5723 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 5769..6197 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5856..5875) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6066..6087 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(6206..6226) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 6211..6540 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 6391..6526 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6453..6472 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 6588..6635 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 6654..7052 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" polyA_signal 7210..7331 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(7247..7266) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 7301..7320 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(7380..7396) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7380..7396) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7393..7415) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7404..7420 /label=lac operator /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-beta-D-thiogalactopyranoside (IPTG)." promoter complement(7428..7458) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7473..7494 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7611..7628) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7782..8370) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7862..7881) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8541..9401) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 9164..9183 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9402..9506) /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 gctggctagt 901 taagcttgcc accatgtccg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat 961 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga 1021 gggcgatgcc acctacggca agctgaccct gaagctgatc tgcaccaccg gcaagctgcc 1081 cgtgccctgg cccaccctcg tgaccaccct gggctacggc ctgcagtgct tcgcccgcta 1141 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca 1201 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt 1261 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg 1321 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc 1381 cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg 1441 cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct 1501 gctgcccgac aaccactacc tgagctacca gtccaaactg agcaaagacc ccaacgagaa 1561 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga 1621 cgagctgtac aagtccggac tcagatctgg aggcttccac agagatgctg atgaaaccaa 1681 agaatggatt gaagagaaga atcaagctct aaacacagac aattatggac atgatctcgc 1741 cagtgtccag gccctgcaac gcaagcatga gggcttcgag agggaccttg cggctctcgg 1801 tgacaaggta aactcccttg gtgaaacagc agagcgcctg atccagtccc atcccgagtc 1861 agcagaagac ctgcaggaaa agtgcacaga gttaaaccag gcctggagca gcctggggaa 1921 acgtgcagat cagcgcaagg caaagggagg cgtgaattcc atggtgagca agggcgagga 1981 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa 2041 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt 2101 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgacctg 2161 gggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc 2221 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta 2281 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa 2341 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaacgccat 2401 cagcgacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa 2461 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac 2521 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccaa 2581 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc 2641 cgccgggatc actctcggca tggacgagct gtacaagatg gaggggtact cagaggaggc 2701 tagcttgctg cggcacctgg agaaggtggc cagtgaggag gaagaggtac cactggtggt 2761 ttatctaaag gagaatgcag cactgctgac agccaatggg ctgcacctgt cccaaaaccg 2821 agaggcccag cagtcctcac cggccccacc tccagctgag gtccacagcc cagctgcaga 2881 tgtcaaccaa aaccttgcct cgcccagtgc cacgctcacc acaccaactt ctaacagcag 2941 ccacaatccg ccagccaccg atgtcaatca gaacccaccg gcaactgttg tcccacagag 3001 cctgccactt tctagcatcc aacagaattc ctcagaggcc caactcccat ctaatggcac 3061 agggcctgct tccaaaccca gcaccctgtg tgctgatggg caaccccagg caccggctga 3121 ggaggtgaga tgcagcacac tcctaattga caaggtatca actccagcta ccaccaccag 3181 caccttctcc agagaagcta cgctcatccc cagctccagg cccccagcct cagatttcat 3241 gtccagctcc ctgctcattg acatccagcc caacacccta gtggtgtcag cagatcaaga 3301 gatgtctggg cgagcagctg ccaccacgcc caccaaggtc tacagtgagg tccacttcac 3361 actggccaag cccccatcag tggtcaacag gacggccagg ccttttggga tccaggcgcc 3421 agggggcacc agccagatgg agaggagccc catgctagag agacgacatt ttggggagaa 3481 ggccccggct ccccagcccc ccagtttgcc agacaggagc ccccggccac agagacacat 3541 aatgtcccgc agccccatgg tggaaaggag gatgatgggg cagcgaagcc cggcctcaga 3601 gagacgcccc ttggggaact tcactgcacc ccccacctac actgagacct tgtccacagc 3661 ccctctggct tcctgggtga ggtctcctcc ctcatattct gtcctgtatc ccagctccga 3721 ccccaagtct tctcatctga agggccaggc ggttcctgcc agcaagacgg gcattctgga 3781 ggagtcgatg gcccgccggg gcagccgcaa atccatgttt actttcgtgg agaagcccaa 3841 ggtgaccccg aatccagact tgctggatct ggtacagaca gcggatgaga agcggcggca 3901 gagggaccag ggggaggtag gcgtggagga ggagcccttc gcactggggg ccgaggcctc 3961 caacttccag caggagccag cacctcgtga cagggccagc cccgcggcgg cggaggaggt 4021 ggtaccagag tgggcctcct gcctcaagtc accccgcatc caggccaagc cgaagcccaa 4081 acccaaccag aacctctccg aggcctctgg gaagggagct gagctctacg cccgccgcca 4141 gtcacggatg gagaaatatg tcatcgagtc ttcaagccac acgccagagc tggcccgctg 4201 cccatcacct accatgtccc tgccttcctc ctggaaatac cccactaacg cccccggggc 4261 cttccgagtg gcatcccgaa gcccagcccg gaccccgcct gcctccctct accatggcta 4321 cctgcctgag aacggggtcc tgcgcccaga gcccaccaag cagccgccat accagctgcg 4381 cccctcgctc tttgtcctct cacctatcaa ggagcctgcc aaggtctcac caagagctgc 4441 ctcgcccgcc aagcccagct ccttggacct ggtgcccaac ctgcccaagg gggctctccc 4501 tccatctcct gccctgcctc ggccctcgcg ctcctcaccg ggcctctaca cctcccccgg 4561 ccaggacagc ctgcagccca ctgccgtgag ccctccttac ggcggtgaca tctcccccgt 4621 gtctccctcc agggcgtggt ctccccgagc caagcaggcc cccaggccct ccttctctac 4681 ccggaacgcc gggatcgagg ctcaggaccg ccgggagagc ctgcccacct ccccaccctg 4741 gacgccgggc gcgtcccggc cccccagcag cctagacggc tgggtgagcc cgggcccgtg 4801 ggagccaggt cgcgggagca gcatgagcag ccccccgccg ctgccgccgc caccgcccat 4861 gtctccctcg tggagcgagc gctcggtgtc cccgctgcga cctgagaccg aggcgcggcc 4921 ccccagccgc cagctgcagg cgcttctggc gcgaaacatc atcaatgcgg cccggcgcaa 4981 gagcgcctcc ccgcggtcgg cgggcgccga gaacccgcgg cccttctccc cgccgagggc 5041 gccaccgccc ccgcccccgc ccccgccccc gcccccgcgc atgcgctcgc cacagcccgc 5101 ccgccccggc tcggctgctg tgccgggggc agccttcgcg cccatcccgc ggagcccgtt 5161 gcccgccggt ccttcgtcct gcaccagtcc ccggagcccg ctgcccgcgc ctcccaggcc 5221 cttcctctac cgccgctcgc ccacggactc cgacgtgtcc ctcgactccg aggactccgg 5281 ggctaagtct ccaggcatcc tgggctacaa tatctgtccc cgcgggtgga atggcagcct 5341 tcggctcaag cgtggcagcc tccccgccga ggcctcctgc accaccctcg agtctagagg 5401 gcccttcgaa caaaaactca tctcagaaga ggatctgaat atgcataccg gtcatcatca 5461 ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc 5521 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt 5581 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct 5641 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc 5701 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg 5761 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag 5821 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt 5881 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt 5941 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg 6001 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt 6061 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt 6121 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca 6181 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca 6241 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt 6301 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 6361 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 6421 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct 6481 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa 6541 aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat 6601 catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca 6661 agcctttgtc tcaagaagaa tccaccctca ttgaaagagc aacggctaca atcaacagca 6721 tccccatctc tgaagactac agcgtcgcca gcgcagctct ctctagcgac ggccgcatct 6781 tcactggtgt caatgtatat cattttactg ggggaccttg tgcagaactc gtggtgctgg 6841 gcactgctgc tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga 6901 acaggggcat cttgagcccc tgcggacggt gccgacaggt gcttctcgat ctgcatcctg 6961 ggatcaaagc catagtgaag gacagtgatg gacagccgac ggcagttggg attcgtgaat 7021 tgctgccctc tggttatgtg tgggagggct aagcacttcg tggccgagga gcaggactga 7081 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc 7141 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc 7201 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 7261 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 7321 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg 7381 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc 7441 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg 7501 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc 7561 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact 7621 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta 7681 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag 7741 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc 7801 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta 7861 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg 7921 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc 7981 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac 8041 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac 8101 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg 8161 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga 8221 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt 8281 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag 8341 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 8401 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 8461 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat 8521 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc 8581 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg 8641 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct 8701 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca 8761 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg 8821 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg 8881 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc 8941 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag 9001 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg 9061 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag 9121 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat 9181 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg 9241 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca 9301 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca 9361 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat 9421 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag 9481 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc //