LOCUS       40924_500        4517 bp DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses LAR-GECO1 in the endoplasmic reticulum in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
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)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4517)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "Biochem J"; date: "2014-11-15- 15"; volume: "464"; issue: "1"; 
            pages: "13-22"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4517
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    complement(42..266)
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        292..310
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             complement(386..1633)
                     /codon_start=1
                     /label=CAR-GECO1
                     /note="CAR-GECO1 is a basic (constitutively fluorescent)
                     red fluorescent protein.  It has high acid sensitivity."
                     /translation="VDSSRRKWNKAGHAWRAIGRLSSPVVSERMYPEDGALKSEIKKGL
                     RLKDGGHYAAEVKTTYKAKKPVQLPGAYVVDIKLDIVSHNEDYTIVEQCERAEGRHSTG
                     GMDELYKGGTGGSLVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEAF
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYIKHPADIPDYFKLSFPEGFRWERVMNFE
                     DGGIIHVNQDSSLQDGVFIYKVKLRGTNFPPDGPVMQKKTMGWEATRDQLTEEQIAEFK
                     EAFSLFDKDGDGTITTKELGTVMRSLGQNPTEAELQDMISEVDADGDGTFDFPEFLTMM
                     ARKMNYTDSEEEIREAFRVADKDGNGYIGAAELRHAMTDIGEKLTDEEVDEMIRVADID
                     GDGQVNYEEFVQMMTAK"
     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"
     enhancer        complement(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
                     /label=AmpR promoter
     CDS             2739..3596
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3770..4358
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
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
//
