LOCUS       sequence-261989-        6944 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Fluorescent calcium reporter and mouse vomeronasal 1 receptor 58.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6944)
  AUTHORS   Lee D, Kume M, Holy TE
  TITLE     Sensory coding mechanisms revealed by optical tagging of 
            physiologically defined neuronal types.
  JOURNAL   Science. 2019 Dec 13;366(6471):1384-1389. doi: 
            10.1126/science.aax8055.
  PUBMED    31831669
REFERENCE   2  (bases 1 to 6944)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2019-12-13- 13"; volume: "366"; 
            issue: "6471"; pages: "1384-1389"
COMMENT     Sequence Label: pAAV2-GCaMP5g-P2A-Vmn1r58
FEATURES             Location/Qualifiers
     source          1..6944
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        223..526
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1050..1069
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1124..1133
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1130..1138,1139..1156,1157..1159,1160..1192,1193..1195,
                     1196..1219,1220..1246,1247..1303,1304..1309,1310..1579,
                     1580..1597,1598..2032,2033..2038,2039..2479)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYTDSEEEIREAFRVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             join(1130..1138,1139..1156,1157..1159,1160..1192,1193..1195,
                     1196..1219,1220..1246,1247..1303,1304..1309,1310..1579,
                     1580..1597,1598..2032,2033..2038,2039..2479)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYTDSEEEIREAFRVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      1226..1235
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1516..1537
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1643..1664)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1904..1923)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2495..2551
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     polyA_signal    3506..3627
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3517..3536)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3571..3590
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3713..3730)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3719..3926
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3956..4096
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3956..4085
                     /label=AAV2 ITR
     rep_origin      4182..4637
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4269..4288)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4479..4500
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4704..4723)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4823..4845
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4883..4901)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4969..5073
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5074..5142,5143..5934)
                     /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(5292..5311)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6105..6693
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6594..6613
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtagatct gaattcggta cctagttatt aatagtaatc
      181 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      241 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      301 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      361 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      421 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      481 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc
      541 cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt
      601 tattttttaa ttattttgtg cagcgatggg ggcggggggg ggcgccaggc ggggcggggc
      661 ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg
      721 cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa
      781 gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg
      841 cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg
      901 acggcccttc tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc
      961 tgtggctgcg tgaaagcctt gaggggctcc gggagctaga gcctctgcta accatgttca
     1021 tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc tgtctcatca
     1081 ttttggcaaa gaattcctcg aagatccgaa ggggttcaag cttgccacca tgggttctca
     1141 tcatcatcat catcatggta tggctagcat gactggtgga cagcaaatgg gtcgggatct
     1201 gtacgacgat gacgataagg atctcgccac catggtcgac tcatcacgtc gtaagtggaa
     1261 taagacaggt cacgcagtca gagctatagg tcggctgagc tcactcgaga acgtctatat
     1321 caaggccgac aagcagaaga acggcatcaa ggcgaacttc aagatccgcc acaacatcga
     1381 ggacggcggc gtgcagctcg cctaccacta ccagcagaac acccccatcg gcgacggccc
     1441 cgtgctgctg cccgacaacc actacctgag cgtgcagtcc aaactttcga aagaccccaa
     1501 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     1561 catggacgag ctgtacaagg gcggtaccgg agggagcatg gtgagcaagg gcgaggagct
     1621 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     1681 cagcgtgtcc ggcgagggtg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     1741 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     1801 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     1861 catgcccgaa ggctacatcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     1921 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     1981 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca acctgccgga
     2041 ccaactgact gaagagcaga tcgcagaatt taaagaggct ttctccctat ttgacaagga
     2101 cggggatggg acaataacaa ccaaggagct ggggacggtg atgcggtctc tggggcagaa
     2161 ccccacagaa gcagagctgc aggacatgat caatgaagta gatgccgacg gtgacggcac
     2221 aatcgacttc cctgagttcc tgacaatgat ggcaagaaaa atgaaataca cagacagtga
     2281 agaagaaatt agagaagcgt tccgtgtgtt tgataaggat ggcaatggct acatcagtgc
     2341 agcagagctt cgccacgtga tgacaaacct tggagagaag ttaacagatg aagaggttga
     2401 tgaaatgatc agggaagcag acatcgatgg ggatggtcag gtaaactacg aagagtttgt
     2461 acaaatgatg acagcgaagg aattcggaag cggagctact aacttcagcc tgctgaagca
     2521 ggctggagac gtggaggaga accctggacc tggatccatg gaaatgttgg ctcttcagat
     2581 ccttttgctt tgccatgttg tggttgggac tgtgggcaac atccttctgt ttgtccataa
     2641 cttctctcca atcttgactg actccagact gaagccaata caggtcattc taatcaactt
     2701 ggctgtggcc aatgcattca tgctccttct gtttgcatat tcatatgacc ttactgatat
     2761 tgttcccagg aagcctccaa ctgacctcaa atgtaaactt gcatacttct ttcacatggt
     2821 ggcacgaggc acaatcatgt gctctacctg tatcctcagt acataccagt ttgtcactct
     2881 tgtccctggt acctgggcta gggtcatgtt cagagaaata tcccccaagg ttgtgagcta
     2941 ttgttgttgc atctgctggt tgttcagtgt cttaaataat gcttacatcc caatgaatgt
     3001 cagtggtcca cagaaatcac acaatgactc tgattctaaa ggcaattcaa tatgctccat
     3061 ctctggggtc agtgtagaca tgaatatctt gcggttttcc catgacatca tattcctaag
     3121 catcatggcc tggaccagtg tctccatggt gattcaccta cacagacacc accagagaat
     3181 gaaccacata cataaggcca atcaaaacaa cagaggccat gctgagacca gagcagccca
     3241 caccatcctg atgctggtgg tcacatttgt gagcttatat attctaaatt gtattactat
     3301 tttatttcac atttcctttg tggaatctcg tctctggttg aggtatgcca caaaactcct
     3361 ggctctaagc ttccccacca tttctccctt tcttttgata tttagggatc gtaagggtca
     3421 ttgttctctg cacatcattg tcagtatggg aatccacgtg acggggggag ccattacaga
     3481 gggacagtaa ggatcccaga catgataaga tacattgatg agtttggaca aaccacaact
     3541 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     3601 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     3661 gttcaggggg aggtgtggga ggttttttag tcgactagag ctcgctgatc agcctcgact
     3721 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     3781 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     3841 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     3901 gaagacaata gcaggcatgc tggggagaga tctcacgtgc ggaccgagcg gccgcaggaa
     3961 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     4021 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     4081 cgcagctgcc tgcaggggcg cctgatgcca gctggcgtaa tagcgaagag gcccgcaccg
     4141 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct gtagcggcgc
     4201 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4261 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4321 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4381 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4441 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4501 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4561 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4621 attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     4681 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     4741 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     4801 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     4861 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     4921 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4981 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5041 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5101 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5161 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5221 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5281 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     5341 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5401 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5461 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5521 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5581 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5641 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5701 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5761 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5821 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5881 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5941 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6001 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6061 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6121 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6181 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6241 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6301 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6361 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6421 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6481 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6541 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6601 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6661 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6721 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6781 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     6841 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     6901 cccgcgcgtt ggccgattca ttaatgcagc tggggctcac atgt
//
