LOCUS Exported 4517 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses LAR-GECO1 in the endoplasmic reticulum in mammalian cells. ACCESSION . VERSION . KEYWORDS CMV-ER-LAR-GECO1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4517) AUTHORS Wu J, Prole DL, Shen Y, Lin Z, Gnanasekaran A, Liu Y, Chen L, Zhou H, Chen SR, Usachev YM, Taylor CW, Campbell RE TITLE Red fluorescent genetically encoded Ca2+ indicators for use in mitochondria and endoplasmic reticulum. JOURNAL Biochem J. 2014 Nov 15;464(1):13-22. doi: 10.1042/BJ20140931. PUBMED 25164254 REFERENCE 2 (bases 1 to 4517) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4517 /organism="synthetic DNA construct" /mol_type="other DNA" polyA_signal 42..266 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind 255..272 /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" promoter 292..310 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 292..309 /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(850..869) /label=mCherry-F /note="mCherry, forward primer" primer_bind complement(1721..1740) /label=T7 /note="T7 promoter, forward primer" promoter complement(1722..1740) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter complement(1785..1988) /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind complement(1814..1834) /label=CMV-F /note="Human CMV immediate early promoter, forward primer" enhancer 1989..2368 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" primer_bind 2540..2559 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 2634..2738 /gene="bla" /label=AmpR promoter CDS 2739..3599 /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 complement(2957..2976) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3770..4358 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4259..4278 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ttccggacta gagccccagc tggttctttc cgcctcagaa gccatagagc ccaccgcatc 61 cccagcatgc ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc 121 ccccagaata gaatgacacc tactcagaca atgcgatgca atttcctcat tttattagga 181 aaggacagtg ggagtggcac cttccagggt caaggaaggc acgggggagg ggcaaacaac 241 agatggctgg caactagaag gcacagtcga ggctgatcag cgagctctag catttaggtg 301 acactataga atagggccct ctagatgcat gctcgagcgg ccgccagtgt gatggatatc 361 tgcagaattc ttacagctcg tccttcttcg ctgtcatcat ttgtacaaac tcttcgtagt 421 ttacctgacc atccccatcg atgtctgcta ccctgatcat ttcatcaacc tcctcatctg 481 ttaacttctc tccaatgtct gtcatcgcgt ggcgaagctc tgctgcgccg atgtagccat 541 tgccgtcctt atccgccacg cggaacgctt ctctaatttc ctcttcactg tctgtgtaat 601 tcatttttct tgccatcatc gtcaggaact cagggaagtc gaatgtgccg tcaccgtcgg 661 catctacttc actgatcatg tcctgcagct ctgcttctgt ggggttctgc cccagagacc 721 gcatcaccgt ccccagctcc ttggttgtta tcgtcccatc cccgtccttg tcaaataggg 781 agaaagcctc tttaaattct gcgatctgct cttcagtcag ttggtcgcgc gtagcctccc 841 agcccatggt cttcttctgc attacggggc cgtcgggggg gaagttggtg ccgcgcagct 901 tcaccttgta gatgaatacg ccgtcctgca gggaggagtc ctggttaacg tgaataatgc 961 cgccgtcctc gaagttcatc acgcgctccc acctgaagcc ctcggggaag gacagcttga 1021 agtagtcggg gatgtcggct gggtgcttaa tgtaggcctt ggagccgtac atgaactgag 1081 gggacaggat gtcccaggcg aagggcaggg ggccaccctt ggtcaccttc agcttagcgg 1141 tctgaaaggc ctcgtagggg cggccctcgc cctcgccctc gatctcgaac tcgtggccgt 1201 tcacggagcc ctccatgtgc accttgaagc gcatgaactc cttgatgatg gccatgttat 1261 cctcctcgcc cttgctcacc agactcccgc ctgtacctcc cttgtacagc tcgtccatgc 1321 cgccggtgga gtggcggccc tcggcgcgtt cgcactgttc cacgatggtg tagtcctcgt 1381 tgtgggacac gatgtccaac ttgatgtcga cgacgtaggc gccgggcagc tgcacgggct 1441 tcttggcctt gtaggtggtc ttgacctcgg cggcgtagtg gccgccgtcc ttcagcctca 1501 gccccttctt gatctcgctc ttcagggctc cgtcctcggg gtacatccgc tcggaaacca 1561 cgggtgagct cagccgacct atagctctcc atgcgtgacc tgccttattc cacttacgac 1621 gtgaagagtc gacgtcggcg gcggcgccca gcaggcccag cagcaggggg acgggcagca 1681 gcatggcggt ggatccgagc tcggtaccaa gcttgggtct ccctatagtg agtcgtatta 1741 atttcgataa gccagtaagc agtgggttct ctagttagcc agagagctct gcttatatag 1801 acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat 1861 tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg 1921 agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac 1981 cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc 2041 cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata 2101 gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa 2161 tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc 2221 attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta 2281 agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac 2341 tattaataac tagtcaataa tcaatgtcaa cgcgtatatc tggcccgtac atcgcgaagc 2401 agcgcaaaac gcctaaccct aagcagattc ttcatgcaat tgtcggtcaa gccttgcctt 2461 gttgtagctt aaattttgct cgcgcactac tcagcgacct ccaacacaca agcagggagc 2521 agatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatagggga 2581 tcgggagatc tcccgatccg tcgacgtcag gtggcacttt tcggggaaat gtgcgcggaa 2641 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac 2701 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg 2761 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc 2821 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg 2881 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga 2941 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc 3001 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag 3061 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga 3121 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg 3181 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga 3241 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt 3301 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact 3361 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt 3421 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg 3481 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta 3541 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac 3601 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta 3661 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt 3721 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt 3781 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt 3841 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc 3901 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg 3961 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg 4021 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt 4081 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac 4141 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg 4201 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg 4261 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat 4321 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt 4381 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg 4441 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa 4501 cgaccgagcg cagcgag //