LOCUS       Exported                5368 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses red fluorescent calcium sensor in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pGP-CMV-NEW-fRGECO2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5368)
  AUTHORS   Kerruth S, Coates C, Durst CD, Oertner TG, Torok K
  TITLE     The kinetic mechanisms of fast-decay red-fluorescent genetically 
            encoded calcium indicators.
  JOURNAL   J Biol Chem. 2019 Mar 15;294(11):3934-3946. doi: 
            10.1074/jbc.RA118.004543. Epub 2019 Jan 16.
  PUBMED    30651353
REFERENCE   2  (bases 1 to 5368)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5368
                     /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"
     CDS             688..705
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             709..741
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             745..768
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     1548..1567
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2156..2277
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2193..2212)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2247..2266
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2284..2739)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2421..2442)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2633..2652
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2766..2870
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2872..3229
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2895..2915)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3080..3215
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3142..3161
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3264..4058
                     /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(3318..3337)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3928..3947
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4246..4265)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4290..4337
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4666..5254
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5155..5174
                     /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 gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgccac catgctgcag aacgagcttg ctcttaagtt ggctggactt
      661 gatattaaca agactggagg aggttctcat catcatcatc atcatggtat ggctagcatg
      721 actggtggac agcaaatggg tcgggatctg tacgacgatg acgataagga tctcgcaaca
      781 atggtcgact catcacgtcg taagtacaat aaggcaggtc acgcagtcag agctataggt
      841 cggctgagct cacccgtggt ttccgagcgg atgtaccccg aggacggcgc cctgaagagc
      901 gagatcaaga aggggctgag gctgaaggac ggcggccact acgccgccga ggtcaagacc
      961 acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acatcgtgga catcaagttg
     1021 gacatcgtgt cccacaacga ggactacacc atcgtggaac agtgcgaacg cgccgagggc
     1081 cgccactcca ccggcggcat ggacgagctg tacaagggag gtacaggcgg gagtctggtg
     1141 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     1201 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     1261 tacgaggcct ttcagaccgc taagctgaag gtgaccaagg gtggccccct gcccttcgcc
     1321 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacattaa gcacccagcc
     1381 gacatccccg actacttcaa gctgtccttc cccgagggct tcaggtggga gcgcgtgatg
     1441 aacttcgagg acggcggcat tattcacgtg aaccaggact cctccctgca ggacggcgta
     1501 ttcatctaca aggtgaagct gcgcggcacc aacttccccc ccgacggccc cgtaatgcag
     1561 aagaagacca tgggctggga ggctacgcgt gacgacctga ctgaagagca gatcgcagaa
     1621 tttaaagagg ctttctccct atttgacaag gacggggatg ggacgataac aaccaaggag
     1681 cttgggacgg tgttccgatc gctggggcag aaccccacag aagcagagct gcaggacatg
     1741 atcaatgaag tagatgccga cggtgacggc acattcgact tccctgagtt cctgacgatg
     1801 atggcaagaa aaatgaatga cacagacagt gaagaggaaa ttcgcgaagc gttccgcgtg
     1861 tttgataagg acggcaatgg ctacatcggc gcagcagagc ttcgccacgt gatgacagac
     1921 cttggagaga agttaacaga tgaggaggtt gatgaaatga tcagggtagc agccatcgat
     1981 ggggatggtc aggtaaacta cgaagagttt gtacaaatga tgacagcgaa gtaggcggcc
     2041 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     2101 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     2161 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     2221 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     2281 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     2341 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     2401 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     2461 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     2521 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     2581 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     2641 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     2701 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     2761 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2821 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     2881 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     2941 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3001 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3061 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3121 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     3181 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     3241 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     3301 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     3361 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     3421 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     3481 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     3541 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     3601 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     3661 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     3721 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     3781 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     3841 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     3901 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     3961 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4021 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     4081 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4141 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4201 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     4261 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     4321 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     4381 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     4441 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     4501 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4561 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4621 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4681 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4741 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4801 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4861 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4921 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4981 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5041 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5101 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5161 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5221 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5281 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     5341 tgtggataac cgtattaccg ccatgcat
//
