LOCUS       sequence-412311-        5934 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control for CybSEP2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5934)
  AUTHORS   Kim D-I, Park S, Ye M, Chen JY, Jhang J, Hunker AC, Zweifel LS, 
            Palmiter RD, Han S
  TITLE     Novel genetically encoded tools for imaging or silencing 
            neuropeptide release from presynaptic terminals in vivo
  JOURNAL   bioRxiv 2023.01.19.524797
REFERENCE   2  (bases 1 to 5934)
  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: "bioRxiv 
            2023.01.19.524797"
COMMENT     Sequence Label: pAAV-hSyn1-FLEX-CybGam-WPRE
FEATURES             Location/Qualifiers
     source          1..5934
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        166..613
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(646..679)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(690..723)
                     /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)."
     CDS             complement(735..758)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(1170..1886)
                     /codon_start=1
                     /product="acid-tolerant monomeric green fluorescent protein
                     (Shinoda et al., 2018)"
                     /label=Gamillus
                     /translation="MVSKGEEASGRALFQYPMTSKIELNGEINGKKFKVAGEGFTPSSG
                     RFNMHAYCTTGDLPMSWVVIASPLQYGFHMFAHYPEDITHFFQECFPGSYTLDRTLRME
                     GDGTLTTHHEYSLEDGCVTSKTTLNASGFDPKGATMTKSFVKQLPNEVKITPHGPNGIR
                     LTSTVLYLKEDGTIQIGTQDCIVTPVGGRKVTQPKAHFLHTQIIQKKDPNDTRDHIVQT
                     ELAVAGNLWHGMDELYK"
     regulatory      2222..2231
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2237..2270
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2281..2314
                     /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)."
     misc_feature    2338..2926
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2391..2411)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2809..2820)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2961..2978)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2967..3174
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3197..3337
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3197..3326
                     /label=AAV2 ITR
     rep_origin      3412..3867
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3499..3518)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3709..3730
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3884..3903)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4003..4025
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4063..4081)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4149..4253
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4254..4322,4323..5114)
                     /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(4472..4491)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5285..5873
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5774..5793
                     /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 gcgtctgcag agggccctgc gtatgagtgc aagtgggttt
      181 taggaccagg atgaggcggg gtgggggtgc ctacctgacg accgaccccg acccactgga
      241 caagcaccca acccccattc cccaaattgc gcatccccta tcagagaggg ggaggggaaa
      301 caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca ccgcggacag tgccttcgcc
      361 cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa ggcgcgctga cgtcactcgc
      421 cggtcccccg caaactcccc ttcccggcca ccttggtcgc gtccgcgccg ccgccggccc
      481 agccggaccg caccacgcga ggcgcgagat aggggggcac gggcgcgacc atctgcgctg
      541 cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc agcggaggag tcgtgtcgtg
      601 cctgagagcg cagacgcgtg gacggtaccc ttcaggtcga cgtcgataac ttcgtatagc
      661 atacattata cgaagttatc cgaatcgcaa taacttcgta taggatactt tatacgaagt
      721 tatttggcgc gtcacttatc gtcgtcatcc ttgtaatcct ggggactggg gctgtctccc
      781 tccgtgagtg tcttgaagtc catggagagg gcttgctctt cagcctggga aggccgcttc
      841 cagtcagcct gagccaagat gtagagcaca accaccccaa aacagaccag cagcaagcct
      901 agcacgttgg ccagcacccc ctcaggttca aacgtgctgt acttggaccc caatttaaac
      961 agcagggcct ccttcaggcc cagtagggct gtgcccacag agaagaggaa gatggtggca
     1021 ccaaagaaaa tggcctgagg gcggtagcgg ctccgcagag agaatgaagc tccagggaac
     1081 aggaagaagc tgaaacccac gagccactgc acaaagtaca gaacaaagac taagatccca
     1141 caccagctgt gcaagctgta caggtctgcc ttgtacagct cgtccatgcc gtgccacaga
     1201 ttgccggcca cggccagttc tgtctgcacg atgtggtccc gggtgtcgtt ggggtccttc
     1261 ttctggatga tctgggtgtg caggaagtgg gccttgggct gggtcacttt ccttccgccc
     1321 acaggggtca cgatgcagtc ctgggtgccg atctggatgg tgccatcctc cttcaggtac
     1381 agcacggtgc tggtcagtct gatgccgttg gggccgtggg gggtgatctt cacctcgtta
     1441 ggcagttgtt tcacgaagct cttggtcatg gtggcgccct tagggtcgaa gccgctggca
     1501 ttcagggtgg tcttggaggt cacgcagccg tcctccaggc tgtactcgtg gtgggtggtc
     1561 agggtgccgt cgccttccat tctcagggtc cggtccaggg tgtagctgcc ggggaagcat
     1621 tcctggaaaa agtgtgtgat gtcctcgggg tagtgggcga acatgtggaa gccgtactgg
     1681 agggggctgg caatcacgac ccagctcata ggcaggtcgc cggtggtgca gtaggcgtgc
     1741 atgttgaatc tgccgctgct gggggtgaag ccctcgccgg ccaccttgaa tttcttgccg
     1801 ttgatctcgc cgttcagctc gatcttgctg gtcatggggt actggaacag ggctctgcca
     1861 gatgcctcct cgcccttgct caccatatag cccttcttct tgtggtagtc gaacacagcc
     1921 accaacccca ccagggcgat gatgaaggcg aagacgtgca gcagcccagc caagatcttg
     1981 gtcgtgcgtt tggcttctct tctgaagaca cggtacacca gcagagcatc tccttgaagg
     2041 aagatcatgc ctatgaccat gcagagcggg tgcacattaa actgcaggga gctttcccag
     2101 gcaatgccac ctcggtacag acccagccac gcaccagtca cagccactac agtgaggccc
     2161 agcagctggg agaaggccac atagtacggc agtgcagcag ggacagacgc agaactgtgc
     2221 tccatggtgg cccttgataa cttcgtataa tgtatgctat acgaagttat ccgaatcgca
     2281 ataacttcgt ataaagtatc ctatacgaag ttatcgaaga tcatcaagct tatcgataat
     2341 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     2401 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     2461 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     2521 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     2581 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     2641 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     2701 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     2761 tatgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     2821 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     2881 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgagcg
     2941 ctgctcgaga gatcgatctg cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3001 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3061 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3121 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggacacgtg
     3181 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     3241 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     3301 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     3361 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     3421 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3481 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3541 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3601 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     3661 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3721 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     3781 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3841 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     3901 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     3961 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     4021 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     4081 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     4141 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4201 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4261 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4321 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4381 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4441 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4501 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4561 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4621 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4681 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4741 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4801 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4861 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4921 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4981 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5041 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5101 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     5161 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5221 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5281 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5341 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5401 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5461 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5521 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5581 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5641 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5701 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5761 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5821 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5881 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
