LOCUS       sequence-261976-        6957 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent calcium reporter and mouse vomeronasal 1 receptor 89.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6957)
  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 6957)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 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-Vmn1r89
FEATURES             Location/Qualifiers
     source          1..6957
                     /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     1051..1070
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1125..1134
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1131..1139,1140..1157,1158..1160,1161..1193,1194..1196,
                     1197..1220,1221..1247,1248..1304,1305..1310,1311..1580,
                     1581..1598,1599..2033,2034..2039,2040..2480)
                     /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(1131..1139,1140..1157,1158..1160,1161..1193,1194..1196,
                     1197..1220,1221..1247,1248..1304,1305..1310,1311..1580,
                     1581..1598,1599..2033,2034..2039,2040..2480)
                     /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      1227..1236
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1517..1538
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1644..1665)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1905..1924)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2496..2552
                     /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    3519..3640
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3530..3549)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3584..3603
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3726..3743)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3732..3939
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3969..4109
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3969..4098
                     /label=AAV2 ITR
     rep_origin      4195..4650
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4282..4301)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4492..4513
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4717..4736)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4836..4858
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4896..4914)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4982..5086
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5087..5155,5156..5947)
                     /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(5305..5324)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6118..6706
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6607..6626
                     /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 gggcgccagg cggggcgggg
      661 cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc
      721 gcgctccgaa agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga
      781 agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc
      841 gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg
      901 gacggccctt ctcctccggg ctgtaattag cgcttggttt aatgacggct tgtttctttt
      961 ctgtggctgc gtgaaagcct tgaggggctc cgggagctag agcctctgct aaccatgttc
     1021 atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc
     1081 attttggcaa agaattcctc gaagatccga aggggttcaa gcttgccacc atgggttctc
     1141 atcatcatca tcatcatggt atggctagca tgactggtgg acagcaaatg ggtcgggatc
     1201 tgtacgacga tgacgataag gatctcgcca ccatggtcga ctcatcacgt cgtaagtgga
     1261 ataagacagg tcacgcagtc agagctatag gtcggctgag ctcactcgag aacgtctata
     1321 tcaaggccga caagcagaag aacggcatca aggcgaactt caagatccgc cacaacatcg
     1381 aggacggcgg cgtgcagctc gcctaccact accagcagaa cacccccatc ggcgacggcc
     1441 ccgtgctgct gcccgacaac cactacctga gcgtgcagtc caaactttcg aaagacccca
     1501 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1561 gcatggacga gctgtacaag ggcggtaccg gagggagcat ggtgagcaag ggcgaggagc
     1621 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     1681 tcagcgtgtc cggcgagggt gagggcgatg ccacctacgg caagctgacc ctgaagttca
     1741 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     1801 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     1861 ccatgcccga aggctacatc caggagcgca ccatcttctt caaggacgac ggcaactaca
     1921 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     1981 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aacctgccgg
     2041 accaactgac tgaagagcag atcgcagaat ttaaagaggc tttctcccta tttgacaagg
     2101 acggggatgg gacaataaca accaaggagc tggggacggt gatgcggtct ctggggcaga
     2161 accccacaga agcagagctg caggacatga tcaatgaagt agatgccgac ggtgacggca
     2221 caatcgactt ccctgagttc ctgacaatga tggcaagaaa aatgaaatac acagacagtg
     2281 aagaagaaat tagagaagcg ttccgtgtgt ttgataagga tggcaatggc tacatcagtg
     2341 cagcagagct tcgccacgtg atgacaaacc ttggagagaa gttaacagat gaagaggttg
     2401 atgaaatgat cagggaagca gacatcgatg gggatggtca ggtaaactac gaagagtttg
     2461 tacaaatgat gacagcgaag gaattcggaa gcggagctac taacttcagc ctgctgaagc
     2521 aggctggaga cgtggaggag aaccctggac ctggatccat gttttcaagt gacacattct
     2581 tccagatctt tctcatattc cagttttgtc ttggtgtgat agggaactca tcactgttaa
     2641 tgttgtatgt atacactttc ttcttcaagc ctcattttaa aaagttggta gattcagttt
     2701 tcatgcacct gacaatagtt aatatgttga caatcgcatt cacattgata aaagatatca
     2761 tgttatcctt tggagtacca aattttctgg atgatatcgg ttgtaagaca gttttgtttt
     2821 cattcagagt cagccggggt ctgtccatct gcaccacctc tgttctaagc acgttccaag
     2881 tcatcaccat cactcccagt aactctaagt gggcctcctg gctcaagcct agactctcta
     2941 catggatttt ttcttcctta ctttgctcct ggcttattaa cctagtcatc tacgggtata
     3001 tgcttgacat ggtaatagcc aaaaccaata aaacccatgt tggcaatgga tattcagatg
     3061 gttactgtca aaacaaacac tttgggaacc acaattcagg gtcatttttt agccttctcc
     3121 tcactcacga tctcttctat gtggctgtca tggtgtgggc cagtctctac atggtgattt
     3181 ttctctacag acactacaag agatcccagc atctccacag tagaagcctt tcctcccagt
     3241 catctcctga gcgcaaagcc actcataaca tcctgtctct ggtgagctgc tttgtgttaa
     3301 tttattgctt gaacaactcc atcacccttt atggctttta tacaaaagaa aaaattccaa
     3361 gattggagtc aattaatgca attttaacaa catgctaccc aactgtctgt ccttttttcc
     3421 tcatgaagaa taataaagtt attttgcaat tcacttcttc attttcggta ctgaggatga
     3481 catgtcatca aagttcactg catggctgac agacatgata agatacattg atgagtttgg
     3541 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     3601 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     3661 ttttatgttt caggttcagg gggaggtgtg ggaggttttt tagtcgacta gagctcgctg
     3721 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3781 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3841 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     3901 gggggaggat tgggaagaca atagcaggca tgctggggag agatctcacg tgcggaccga
     3961 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     4021 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     4081 gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg ccagctggcg taatagcgaa
     4141 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc
     4201 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4261 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4321 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     4381 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     4441 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4501 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     4561 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4621 attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct
     4681 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
     4741 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     4801 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     4861 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     4921 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     4981 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     5041 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     5101 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5161 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5221 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5281 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     5341 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5401 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5461 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     5521 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5581 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     5641 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     5701 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5761 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     5821 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5881 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5941 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     6001 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6061 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6121 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6181 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6241 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6301 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6361 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6421 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6481 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6541 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6601 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6661 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6721 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6781 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     6841 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     6901 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggggct cacatgt
//
