LOCUS       sequence-496828-        7328 bp    DNA     circular SYN 09-JAN-2026
DEFINITION  Cre-dependent expression of synaptophysin-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7328)
  AUTHORS   Surmeli G, Marcu DC, McClure C, Garden DLF, Pastoll H, Nolan MF
  TITLE     Molecularly Defined Circuitry Reveals Input-Output Segregation in 
            Deep Layers of the Medial Entorhinal Cortex.
  JOURNAL   Neuron. 2016 Nov 23;92(4):929. doi: 10.1016/j.neuron.2016.11.011.
  PUBMED    27883905
REFERENCE   2  (bases 1 to 7328)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 9, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Neuron."; 
            date: "2016-11-23"; volume: "92(4):929. doi"; pages: " 
            10.1016/j.neuron.2016.11.011"
COMMENT     Sequence Label: pAAV FLEX Synaptophysin GFP
FEATURES             Location/Qualifiers
     source          1..7328
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        28..46
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     28..45
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     repeat_region   67..196
                     /label=AAV2 ITR
     promoter        502..778
                     /label=chicken beta-actin promoter
     primer_bind     1057..1079
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1141..1160
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1195..1228
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    1291..1324
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(1358..2071,2072..2074,2075..2077))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(1361..2071,2072..2074,2075..2077))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1403..1424)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1752..1771
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2011..2032
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2074..2083
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3038..3071)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(3142..3175)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3214..3802
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3267..3287)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3685..3696)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3829..3846)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3835..4059
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(4269..4598)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4283..4418
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4583..4603
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(4844..5432)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4924..4943)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5603..6394,6395..6463))
                     /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     6226..6245
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6464..6568)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6636..6654
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6692..6714)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6814..6833
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7047..7066
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 cataacctta tgtatcatac acatacgatt taggtgacac tatagaatac acggaattaa
       61 ttctagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg
      121 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
      181 atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc tacgtagcca
      241 tgctctaggt accattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      301 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      361 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      421 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      481 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      541 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      601 tgggggcggg gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg
      661 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
      721 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
      781 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
      841 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      901 gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag
      961 ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggc tgtccgcggg
     1021 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg
     1081 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg
     1141 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattggat ccccataact
     1201 tcgtataaag tatcctatac gaagttatat caaaatagga agaccaatgc ttcaccatcg
     1261 acccgaattg ccaagcatca ccatcgaccc ataacttcgt ataatgtatg ctatacgaag
     1321 ttatgaattc ctgcaggagc tcactagtgc ggccgcttta cttgtacagc tcgtccatgc
     1381 cgagagtgat cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt
     1441 tggggtcttt gctcagggcg gactgggtgc tcaggtagtg gttgtcgggc agcagcacgg
     1501 ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc gagctgcacg ctgccgtcct
     1561 cgatgttgtg gcggatcttg aagttcacct tgatgccgtt cttctgcttg tcggccatga
     1621 tatagacgtt gtggctgttg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct
     1681 ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca
     1741 cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt
     1801 agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggc
     1861 tgaagcactg cacgccgtag gtcagggtgg tcacgagggt gggccagggc acgggcagct
     1921 tgccggtggt gcagatgaac ttcagggtca gcttgccgta ggtggcatcg ccctcgccct
     1981 cgccggacac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca
     2041 ccaccccggt gaacagctcc tcgcccttgc tcaccatggt ggcgatggat cccatctgat
     2101 tggagaagga ggtgggcgca ccctgttggc catagccttg ctgcccatag tcgccctgag
     2161 gcccgtagcc accgccaccg ctggctggct gcccgtaatc gggttgataa ccaccctgag
     2221 gcccgtagga gtcctggggc ccatagcctc cggggccctg cccgtagccc gcatcgccgt
     2281 aggcatcgcc aggtgctggt tgcttttccg gggcgcctgg aggtgcgcgc atgaatgggg
     2341 ctgcccagcc tgtctccttg aacacgaacc ataagttgcc aacccagagc accaggttca
     2401 ggaagccaaa caccactgag gtgttgagtc ctgaagtcac agggtccctc agttccttgc
     2461 atgtgttccc tgtctggcgg cacatgggca tctccttgat aatgttctct gggtccgtgg
     2521 ccatcttcac atcggacagg cctttggccc aggctgatga actaactagc cacatgaaag
     2581 cgaacacggc tgtagccaga aagtccatca taggcccttt gttgttctct cggtacttgt
     2641 tctgcaggaa gatgtaggtg gccagggccc ccatggagta gaggaaggca aacacagcca
     2701 cggtgacaaa gaattcagcc gacgaggagt agtccccaac caggaagatc ttggtagtgc
     2761 cccctttgac gcaggagggt gcatcaaagt acacttggtg cagcctgaag gggtactcga
     2821 attcaacttc gatgttgagg gcactctccg tcttgttggc acactccacg ctcagccgaa
     2881 gctccccggt gtagctgcca cacgtagcaa aggcgaagat ggcaaagacc cactgcagca
     2941 ccttcacgaa gccaaggggc tccttgacca cccggaactg acccccagcc accagctgat
     3001 tcaccacgtc cattgctgct gccccgaatt cgatatcata acttcgtata ggatacttta
     3061 tacgaagtta tcattgggat tcttcctatt ttgatccaag catcaccatc gaccctctag
     3121 tccagatctc accatcgacc cataacttcg tatagcatac attatacgaa gttatgtccc
     3181 tcgaagaggt tcaagcttta acagcttatc gataatcaac ctctggatta caaaatttgt
     3241 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     3301 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     3361 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     3421 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     3481 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     3541 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     3601 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     3661 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     3721 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     3781 tccctttggg ccgcctcccc gcatcgatac cgtcgactcg ctgatcagcc tcgactgtgc
     3841 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     3901 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     3961 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     4021 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     4081 gctggggctc gactagagca tggctacgta gataagtagc atggcgggtt aatcattaac
     4141 tacaaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     4201 gaggccgggc gaccaaaggt cgcccgacgc ccgggcggcc tcagtgagcg agcgagcgcg
     4261 cagagctttt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt ctggaatagc
     4321 tcagaggccg aggcggcctc ggcctctgca taaataaaaa aaattagtca gccatggggc
     4381 ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag gggcgggact
     4441 atggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg
     4501 gactttccac acctggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct
     4561 ggggagcctg gggactttcc acaccctaac tgacacacat tccacagctg cattaatgaa
     4621 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4681 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4741 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4801 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4861 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4921 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4981 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5041 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5101 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5161 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5221 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5281 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5341 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5401 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5461 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5521 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5581 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5641 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5701 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5761 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5821 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5881 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5941 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6001 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6061 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6121 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6181 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6241 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6301 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6361 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6421 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6481 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6541 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     6601 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     6661 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     6721 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     6781 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     6841 tgcaccattc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg
     6901 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac
     6961 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     7021 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     7081 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatctg gctagcgatg
     7141 accctgctga ttggttcgct gaccatttcc gggtgcggga cggcgttacc agaaactcag
     7201 aaggttcgtc caaccaaacc gactctgacg gcagtttacg agagagatga tagggtctgc
     7261 ttcagtaagc cagatgctac acaattaggc ttgtacatat tgtcgttaga acgcggctac
     7321 aattaata
//
