LOCUS       sequence-308834-        7302 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human synapsin-1 promoter; Flp-dependent KORD-IRES-mCitrine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7302)
  TITLE     Unpublished
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7302)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-hSyn-fDIO-HA-KORD-IRES-mCitrine-WPREpA
FEATURES             Location/Qualifiers
     source          1..7302
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        147..594
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    632..665
                     /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(764..1474,1475..1477,1478..1480))
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(806..827)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1155..1174
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1414..1435
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1477..1486
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1487..2060
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1647..1666)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     1876..1893
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             2574..2585
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             complement(3219..3245)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(3259..3292)
                     /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    3401..3989
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3454..3474)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3872..3883)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4021..4497
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4220..4239)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4537..4677
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4537..4666
                     /label=AAV2 ITR
     rep_origin      4752..5207
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4839..4858)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5049..5070
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5224..5243)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5343..5365
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5403..5421)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5489..5593
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5594..5662,5663..6454)
                     /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(5812..5831)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6625..7213
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7114..7133
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(7275..7302,1..102)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag
       61 cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc acgcgtgtgt
      121 ctagactgca gagggccctg cgtatgagtg caagtgggtt ttaggaccag gatgaggcgg
      181 ggtgggggtg cctacctgac gaccgacccc gacccactgg acaagcaccc aacccccatt
      241 ccccaaattg cgcatcccct atcagagagg gggaggggaa acaggatgcg gcgaggcgcg
      301 tgcgcactgc cagcttcagc accgcggaca gtgccttcgc ccccgcctgg cggcgcgcgc
      361 caccgccgcc tcagcactga aggcgcgctg acgtcactcg ccggtccccc gcaaactccc
      421 cttcccggcc accttggtcg cgtccgcgcc gccgccggcc cagccggacc gcaccacgcg
      481 aggcgcgaga taggggggca cgggcgcgac catctgcgct gcggcgccgg cgactcagcg
      541 ctgcctcagt ctgcggtggg cagcggagga gtcgtgtcgt gcctgagagc gcagtcgaga
      601 aggtaccgga tcctctagag tcgactccgg agaagttcct attctctaga aagtatagga
      661 acttcgcaga atggtagctg gattgtagct gctattagca atatgaaacc tcttagaagt
      721 tcctattctt caaaaggtat aggaacttcg gcgcgcccta tcacttgtac agctcgtcca
      781 tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct
      841 cgttggggtc tttgctcagt ttggactggt agctcaggta gtggttgtcg ggcagcagca
      901 cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt
      961 cctcgatgtt gtggcggatc ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca
     1021 tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt
     1081 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
     1141 tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga
     1201 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
     1261 gggcgaagca catcaggccg tagccgaagg tggtcacgag ggtgggccag ggcacgggca
     1321 gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc
     1381 cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg
     1441 gcaccacccc ggtgaacagc tcctcgccct tgctcaccat ggtggcttat catcgtgttt
     1501 ttcaaaggaa aaccacgtcc ccgtggttcg gggggcctag acgttttttt aacctcgact
     1561 aaacacatgt aaagcatgtg caccgaggcc ccagatcaga tcccatacaa tggggtacct
     1621 tctgggcatc cttcagcccc ttgttgaata cgcttgagga gagccatttg actctttcca
     1681 caactatcca actcacaacg tggcactggg gttgtgccgc ctttgcaggt gtatcttata
     1741 cacgtggctt ttggccgcag aggcacctgt cgccaggtgg ggggttccgc tgcctgcaaa
     1801 gggtcgctac agacgttgtt tgtcttcaag aagcttccag aggaactgct tccttcacga
     1861 cattcaacag accttgcatt cctttggcga gaggggaaag acccctagga atgctcgtca
     1921 agaagacagg gccaggtttc cgggccctca cattgccaaa agacggcaat atggtggaaa
     1981 ataacatata gacaaacgca caccggcctt attccaagcg gcttcggcca gtaacgttag
     2041 ggggggggga gggagagggg cgatatcgcg gccgctcata ctggtttatt catcccatcg
     2101 atgtccctca ggtaagcagg atcctgaact gtatttcgga ctctgctagt gctctgccgc
     2161 tccatcctca tcttcagtgg aaagcagaag tcccggaaac accgcttgaa gttttcatca
     2221 agaaaggcgt agagaatggg attcaggcta ctgttggtat agcctaaggc gatgcagaag
     2281 taatagctgg agagagcagc tgtgctgtgg gaggtgctcc ccagagcctc caccaggatg
     2341 aatatgtgaa tgggagtcca gcagacgacg aagactgcca ccaccaccag gaccagtctg
     2401 gtgatcctac gcaggttgcg atctttctct cgggagccag aaaggagccg gacgctcttg
     2461 agacgcagga tcatcagggt gtagcagacg atgatgatga ggacagggat cacgaaggca
     2521 aagatgaaga cgcagatctt catgaagagg tcccaccagg agtagtcatc atctgggaac
     2581 tgcaaggagc actcaatgac atcgacgtct tccctgactt tggtgcctcc aaggactatt
     2641 gcagagatgc caacagatga cgacagcagc cagatgcaga tattgatgat ctttgccttc
     2701 aagggtgtgc ggaagtccaa agccttcacg gggtggcaca cggcaatgta gcggtccacg
     2761 ctcatcatgg tcaaggtgaa gatgctggtg aacatgttgt agtaattaat ggaaattact
     2821 atcttgcaca gcacatcccc aaaaggccag gaattcatca agtagaccgt actctgaaag
     2881 ggcatggttg tagtaactaa agcatctgcc aaagccaggt taaatatgta aatgttggtt
     2941 gctgtcttca tctttgtgta tcggatgatc acgaacatga ccagcgagtt gcccaccaag
     3001 cccacgacga acactacgga gtagaccgcc gtgatgatga ccgggatggc cggggagatg
     3061 tgcgcgggct ccagctgcgc gtcctccgag ccggcgctgc cgttgctgtc gggctcggcc
     3121 cagccgggaa accaggcgct gctgttgggg ggcaggcagg cgctcggggc gcaggtaggg
     3181 cccggctccc cgcggaagat ctgaatcggg gattccatag cgtaatctgg aacatcgtat
     3241 gggtacatgg tggctagcga agttcctata ctttctagag aataggaact tcttgcctta
     3301 acccagaaat tatcactgtt attctttaga atggtgcaaa gagaagttcc tatacctttt
     3361 gaagaatagg aacttcgaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa
     3421 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3481 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3541 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     3601 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3661 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     3721 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     3781 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg ccacctggat
     3841 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3901 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3961 tcggatctcc ctttgggccg cctccccgca tcgataccga gcgctgctcg agagatctac
     4021 gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca
     4081 gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc
     4141 ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca
     4201 acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg
     4261 ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg
     4321 ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg
     4381 ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct
     4441 tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg
     4501 attttgtagg taaccacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg
     4561 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     4621 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     4681 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     4741 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4801 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
     4861 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4921 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     4981 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     5041 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
     5101 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
     5161 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     5221 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     5281 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     5341 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     5401 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     5461 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     5521 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     5581 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     5641 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     5701 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     5761 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     5821 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     5881 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     5941 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     6001 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     6061 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     6121 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     6181 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     6241 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     6301 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     6361 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     6421 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     6481 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     6541 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6601 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6661 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6721 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     6781 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6841 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6901 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6961 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     7021 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     7081 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     7141 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     7201 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     7261 ttttgctcac atgtcctgca ggcagctgcg cgctcgctcg ct
//
