LOCUS       sequence-337637-        6971 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Flp-dependent expression of jGCaMP7b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6971)
  AUTHORS   Bauer J, Weiler S, Fernholz MHP, Laubender D, Scheuss V, Hubener M, 
            Bonhoeffer T, Rose T
  TITLE     Limited functional convergence of eye-specific inputs in the 
            retinogeniculate pathway of the mouse.
  JOURNAL   Neuron. 2021 Aug 4;109(15):2457-2468.e12. doi: 
            10.1016/j.neuron.2021.05.036. Epub 2021 Jun 18.
  PUBMED    34146468
REFERENCE   2  (bases 1 to 6971)
  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: 
            "10.1016/j.neuron.2021.05.036"; journalName: "Neuron"; date: 
            "2021-08-4- 4"; volume: "109"; issue: "15"; pages: "2457-2468.e12"
COMMENT     Sequence Label: pAAV1-EF1-alpha-F-Flex-jGCaMP7b-WPR
FEATURES             Location/Qualifiers
     source          1..6971
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   50..179
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        280..1458
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          511..1449
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1406..1426
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1549..1582
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             complement(join(1604..2047,2048..2053,2054..2488,2489..2506,
                     2507..2776,2777..2782,2783..2839,2840..2866,2867..2890,
                     2891..2893,2894..2926,2927..2929,2930..2947,2948..2956))
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNPPDQLTEEQIAEFKELFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMYARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     primer_bind     2163..2182
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2422..2443
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2549..2570)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      2851..2860
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2953..2962
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3087..3120)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    3147..3735
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3200..3220)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3618..3629)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3767..4243
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3966..3985)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4283..4423
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4283..4412
                     /label=AAV2 ITR
     rep_origin      4498..4953
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4585..4604)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4795..4816
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4970..4989)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5089..5111
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5149..5167)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5235..5339
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5340..5408,5409..6200)
                     /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(5558..5577)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6371..6959
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6860..6879
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtc ctgcaggcag
       61 ctgcgcgctc gctcgctcac tgaggccgcc cgggcgtcgg gcgacctttg gtcgcccggc
      121 ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctg
      181 cggccgcacg cgtaagcttt gcaaagatgg ataaagtttt aaacagagag gaatctttgc
      241 agctaatgga ccttctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg
      301 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc
      361 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
      421 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
      481 tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct
      541 ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg gctgcagtac
      601 gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct
      661 taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc
      721 gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt
      781 taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg
      841 ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt
      901 gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga
      961 cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc
     1021 gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg
     1081 ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg
     1141 gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt
     1201 gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt
     1261 acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg
     1321 gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt
     1381 tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt
     1441 ccatttcagg tgtcgtgagg taccgaagtt cctattctat cagaagtata ggaacttcgc
     1501 agaatggtag ctggattgta gctgctatta gcaatatgaa acctcttaga agttcctatt
     1561 ctctagaaag tataggaact tcgaattctt acttgtacag ctttcacttc gctgtcatca
     1621 tttgtacaaa ctcttcgtag tttacctgac catccccatc gatgtctgct tccctgatca
     1681 tttcatcaac ctcttcatct gttaacttct ctccaaggtt tgtcatcacg tggcgaagct
     1741 ctgctgcact gatgtagcca ttgccatcct tatcaaacac accgaacgct tctctaattt
     1801 cttcttccgt gtccctgtat ttcatttttc ttgcgtacat tgtcaggaac tcagggaagt
     1861 cgattgtgcc gtcaccgtcg gcatctactt cattgatcat gtcctgcagc tctgcttctg
     1921 tggggttctg ccccagagac cgcatcaccg tccccagctc cttggttgtt attgtcccat
     1981 ccccgtcctt gtcaaatagg gagaaaagct ctttaaattc tgcgatctgc tcttcagtca
     2041 gttggtcagg tgggttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt
     2101 cgatgccctt aagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc
     2161 gggtcttgta gttgccgtcg tccttgaaga agatggtgcg ctcctggatg tagccttcgg
     2221 gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg ctgaagcact
     2281 gcacgccgta ggtcagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg
     2341 tgcagatgaa cttcagggtc agcttgccgt aggtggcatc gccctcaccc tcgccggaca
     2401 cgctgaactt gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg
     2461 tgaacagctc ctcgcccttg ctcaccatgc tccctccggt accgcccttg tacagctcgt
     2521 ccatgccgag agtgatcccg gcggcggtca cgaactccag caggaccatg tgatcgcgct
     2581 tctcgttggg gtctttcgaa agtttggact gcacgctcag gtagtggttg tcgggcagca
     2641 gcacggggcc gtcgccgatg ggggtgttct gctggtagtg gtaggcgagc tgcacgccgc
     2701 cgtcctcgat gttgtggcgg atcttgaagt tcgccttgat gccgttcttc tgcttgtcgg
     2761 ccttgatata gacgttctcg agtgagctca gccgacctat agctctgact gcgtgacctg
     2821 tcttattcca cttacgacgt gatgagtcga ccatggtggc gagatcctta tcgtcatcgt
     2881 cgtacagatc ccgacccatt tgctgtccac cagtcatgct agccatacca tgatgatgat
     2941 gatgatgaga acccatggtg gcgggtggat cccggctagc tacgtaggcg cgccttaatt
     3001 aagaagttcc tatacttctg atagaatagg aacttcttgc cttaacccag aaattatcac
     3061 tgttattctt tagaatggtg caaagagaag ttcctatact ttctagagaa taggaacttc
     3121 cagaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
     3181 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     3241 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     3301 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     3361 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     3421 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     3481 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     3541 aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt
     3601 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     3661 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     3721 gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctacgggt ggcatccctg
     3781 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct
     3841 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat
     3901 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg
     3961 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc
     4021 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc
     4081 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt
     4141 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt
     4201 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt tgtaggtaac
     4261 cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     4321 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     4381 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     4441 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     4501 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4561 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4621 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4681 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     4741 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4801 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     4861 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4921 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     4981 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     5041 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     5101 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     5161 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     5221 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5281 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     5341 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5401 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5461 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5521 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5581 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5641 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5701 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5761 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5821 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5881 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5941 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     6001 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     6061 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     6121 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     6181 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     6241 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     6301 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6361 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6421 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6481 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6541 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6601 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6661 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6721 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6781 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6841 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6901 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6961 acgccagcaa c
//
