LOCUS       sequence-273137-        7400 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent expression of synaptophysin-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7400)
  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 7400)
  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"; 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..7400
                     /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
     gap             643..653
                     /estimated_length=11
     primer_bind     1129..1151
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1213..1232
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1267..1300
                     /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    1363..1396
                     /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(1430..2143,2144..2146,2147..2149))
                     /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(1433..2143,2144..2146,2147..2149))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1475..1496)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1824..1843
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2083..2104
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2146..2155
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3110..3143)
                     /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(3214..3247)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3286..3874
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3339..3359)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3757..3768)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3901..3918)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3907..4131
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(4341..4670)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4355..4490
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4655..4675
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(4916..5504)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4996..5015)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5675..6466,6467..6535))
                     /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     6298..6317
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6536..6640)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6708..6726
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6764..6786)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6886..6905
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7119..7138
                     /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 gggsgssgss cggggggggg ggnnnnnnnn nnnggggggg
      661 gngggggggg gggggggggg gggggggggg ggggngnggg gsgggggggg gggggggggg
      721 cggggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct
      781 ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc
      841 gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc
      901 gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg
      961 cccttctcct ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg
     1021 gctgcgtgaa agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg
     1081 gctgtccgcg gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct
     1141 ggcgtgtgac cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc
     1201 tacagctcct gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaattgg
     1261 atccccataa cttcgtataa agtatcctat acgaagttat atcaaaatag gaagaccaat
     1321 gcttcaccat cgacccgaat tgccaagcat caccatcgac ccataacttc gtataatgta
     1381 tgctatacga agttatgaat tcctgcagga gctcactagt gcggccgctt tacttgtaca
     1441 gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat
     1501 cgcgcttctc gttggggtct ttgctcaggg cggactgggt gctcaggtag tggttgtcgg
     1561 gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca
     1621 cgctgccgtc ctcgatgttg tggcggatct tgaagttcac cttgatgccg ttcttctgct
     1681 tgtcggccat gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga
     1741 tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc
     1801 cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc
     1861 gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt
     1921 cggggtagcg gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg
     1981 gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat
     2041 cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga
     2101 ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcaccatg gtggcgatgg
     2161 atcccatctg attggagaag gaggtgggcg caccctgttg gccatagcct tgctgcccat
     2221 agtcgccctg aggcccgtag ccaccgccac cgctggctgg ctgcccgtaa tcgggttgat
     2281 aaccaccctg aggcccgtag gagtcctggg gcccatagcc tccggggccc tgcccgtagc
     2341 ccgcatcgcc gtaggcatcg ccaggtgctg gttgcttttc cggggcgcct ggaggtgcgc
     2401 gcatgaatgg ggctgcccag cctgtctcct tgaacacgaa ccataagttg ccaacccaga
     2461 gcaccaggtt caggaagcca aacaccactg aggtgttgag tcctgaagtc acagggtccc
     2521 tcagttcctt gcatgtgttc cctgtctggc ggcacatggg catctccttg ataatgttct
     2581 ctgggtccgt ggccatcttc acatcggaca ggcctttggc ccaggctgat gaactaacta
     2641 gccacatgaa agcgaacacg gctgtagcca gaaagtccat cataggccct ttgttgttct
     2701 ctcggtactt gttctgcagg aagatgtagg tggccagggc ccccatggag tagaggaagg
     2761 caaacacagc cacggtgaca aagaattcag ccgacgagga gtagtcccca accaggaaga
     2821 tcttggtagt gccccctttg acgcaggagg gtgcatcaaa gtacacttgg tgcagcctga
     2881 aggggtactc gaattcaact tcgatgttga gggcactctc cgtcttgttg gcacactcca
     2941 cgctcagccg aagctccccg gtgtagctgc cacacgtagc aaaggcgaag atggcaaaga
     3001 cccactgcag caccttcacg aagccaaggg gctccttgac cacccggaac tgacccccag
     3061 ccaccagctg attcaccacg tccattgctg ctgccccgaa ttcgatatca taacttcgta
     3121 taggatactt tatacgaagt tatcattggg attcttccta ttttgatcca agcatcacca
     3181 tcgaccctct agtccagatc tcaccatcga cccataactt cgtatagcat acattatacg
     3241 aagttatgtc cctcgaagag gttcaagctt taacagctta tcgataatca acctctggat
     3301 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     3361 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     3421 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     3481 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     3541 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     3601 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     3661 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
     3721 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     3781 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     3841 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgact cgctgatcag
     3901 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     3961 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     4021 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     4081 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     4141 cggaaagaac cagctggggc tcgactagag catggctacg tagataagta gcatggcggg
     4201 ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     4261 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggcgg cctcagtgag
     4321 cgagcgagcg cgcagagctt tttgcaaaag cctaggcctc caaaaaagcc tcctcactac
     4381 ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa aaaaattagt
     4441 cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt
     4501 aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct
     4561 ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata
     4621 cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac attccacagc
     4681 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     4741 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     4801 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4861 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4921 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4981 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5041 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5101 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5161 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5221 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5281 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5341 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5401 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5461 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     5521 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5581 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     5641 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     5701 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     5761 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     5821 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     5881 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     5941 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6001 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6061 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6121 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6181 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6241 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6301 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6361 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6421 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     6481 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     6541 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     6601 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     6661 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     6721 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     6781 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     6841 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     6901 ttgtactgag agtgcaccat tcgacgctct cccttatgcg actcctgcat taggaagcag
     6961 cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag
     7021 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
     7081 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg
     7141 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc
     7201 tggctagcga tgaccctgct gattggttcg ctgaccattt ccgggtgcgg gacggcgtta
     7261 ccagaaactc agaaggttcg tccaaccaaa ccgactctga cggcagttta cgagagagat
     7321 gatagggtct gcttcagtaa gccagatgct acacaattag gcttgtacat attgtcgtta
     7381 gaacgcggct acaattaata
//
