LOCUS Exported 6052 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses sfCherry(1-10) in mammalian cells. ACCESSION . VERSION . KEYWORDS pcDNA3.1-sfCherry(1-10) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6052) AUTHORS Kamiyama D, Sekine S, Barsi-Rhyne B, Hu J, Chen B, Gilbert LA, Ishikawa H, Leonetti MD, Marshall WF, Weissman JS, Huang B TITLE Versatile protein tagging in cells with split fluorescent protein. JOURNAL Nat Commun. 2016 Mar 18;7:11046. doi: 10.1038/ncomms11046. PUBMED 26988139 REFERENCE 2 (bases 1 to 6052) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6052 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 381..584 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 535..555 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 629..648 /label=T7 /note="T7 promoter, forward primer" promoter 629..647 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(822..840) /label=mCherry-R /note="mCherry, reverse primer" primer_bind complement(1409..1426) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1415..1639 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 1685..2113 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1772..1791) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1982..2003 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2122..2142) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2127..2456 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2307..2442 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2369..2388 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2523..3317 /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 GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(2577..2596) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3187..3206 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3491..3612 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3528..3547) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3582..3601 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3661..3677) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3661..3677) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3674..3696) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3685..3701 /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(3709..3739) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3754..3775 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3892..3909) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4063..4651) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4143..4162) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4822..5682) /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 5445..5464 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5683..5787) /gene="bla" /label=AmpR promoter primer_bind complement(5862..5881) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag 601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg 661 gctagcgttt aaacttaagc ttggtaccat ggaggaggac aacatggcca tcatcaagga 721 gttcatgaga ttcaaggtgc acatggaggg cagcgtgaac ggccacgagt tcgagatcga 781 gggcgagggc gagggccacc cctacgaggg cacccagacc gccaagctga aggtgaccaa 841 gggcggcccc ctgcccttcg cctgggacat cctgagcccc cagttcatgt acggcagcaa 901 ggcctacgtg aagcaccccg ccgacatccc cgactacctg aagctgagct tccccgaggg 961 cttcacctgg gagagagtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga 1021 cagcagcctg caggacggcg agttcatcta caaggtgaag ctgctgggca ccaacttccc 1081 cagcgacggc cccgtgatgc agaagaagac catgggctgg gaggccagca ccgagagaat 1141 gtaccccgag gacggcgccc tgaagggcga gatcaaccag agactgaagc tgaaggacgg 1201 cggccactac gacgccgagg tgaagaccac ctacaaggcc aagaagcccg tgcagctgcc 1261 cggcgcctac aacgtggaca tcaagctgga catcaccagc cacaacgagg actgaggatc 1321 cactagtcca gtgtggtgga attctgcaga tatccagcac agtggcggcc gctcgagtct 1381 agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct 1441 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt 1501 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg 1561 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg 1621 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat 1681 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg 1741 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc 1801 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga 1861 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt 1921 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat 1981 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat 2041 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa 2101 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct 2161 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa 2221 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 2281 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt 2341 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct 2401 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc 2461 tcccgggagc ttatatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc 2521 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat 2581 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt 2641 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac 2701 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg 2761 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc 2821 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa 2881 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc 2941 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg 3001 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg 3061 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa 3121 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg 3181 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct 3241 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc 3301 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa 3361 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat 3421 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt 3481 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 3541 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 3601 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg 3661 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 3721 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 3781 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 3841 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 3901 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 3961 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 4021 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 4081 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 4141 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 4201 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 4261 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 4321 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 4381 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 4441 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 4501 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 4561 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 4621 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 4681 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 4741 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 4801 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 4861 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 4921 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 4981 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 5041 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 5101 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 5161 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 5221 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 5281 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 5341 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 5401 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 5461 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 5521 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 5581 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 5641 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 5701 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 5761 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga 5821 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc 5881 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg 5941 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt 6001 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg tt //