LOCUS       sequence-490989-        7987 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for imaging of synaptic vesicle recycling in 
            Dopaminergic Neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7987)
  TITLE     Neuromodulator control of energy reserves in dopaminergic neurons
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7987)
  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            "Unpublished"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-THP-Synaptophysin-pHluorin
FEATURES             Location/Qualifiers
     source          1..7987
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   6..135
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     primer_bind     155..171
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     155..171
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     regulatory      2798..2807
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3353..3362
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3359..4075
                     /codon_start=1
                     /product="pH-sensitive mutant of green fluorescent protein 
                     (Ng et al., 2002)"
                     /label=superecliptic pHluorin
                     /note="excited strongly by blue light at neutral pH but 
                     minimally fluorescent at acidic pH"
                     /translation="MGSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNDHQVYIMADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLFTTSTLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(3392..3420)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4015..4035
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4485..4502)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4491..4715
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    4739..5215
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4938..4957)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5255..5395
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5255..5384
                     /label=AAV2 ITR
     rep_origin      5470..5925
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5557..5576)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5767..5788
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5942..5961)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6061..6083
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6121..6139)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6207..6311
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6312..6380,6381..7172)
                     /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(6530..6549)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7343..7931
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7832..7851
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga
       61 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
      121 tcactagggg ttcctgcggc cgcacgcggg ccgctctaga actagtggat cccccaggcc
      181 tcttgggatt cttctcatca aaaagggcat gggacacagc taaagcgtcc agggctcctc
      241 tgtgcccaca gatgccttag atcttggcac aatgtagtca gccagctccg tgtgtgtgtg
      301 tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtttgca
      361 tgtatctcac agacagtgca caatggcctg gatgtgaaca gaggcaagag tctgggccag
      421 cagttgtctc ccaggagggt ccaaagacat cgtattttca agtttaggcc aggtgctcac
      481 ttgggtgagc tcagacacag acaaaggtct ggagagcaca cattccccac ccccacccag
      541 cttctatgca agcacctcca gctgagacaa gaaaacgaat taaaaagcaa tatttgtgtc
      601 agcgtaagac atttgccgaa aggttaaatc cacactcgtg gtgctgcaca gcagccccct
      661 gtgcaggatt tgttaggcac agctccctcc taccccgtgc cacctgagca aatgccaggc
      721 tgggtgggct ggaaccagcc tgggcttgcc tcaccctgga atccccagca ccctccaaag
      781 gaggaccctg ggagtgggca tagacgccct tcaggtgtgg gcaacagccc ccagtcctca
      841 ggatgaaagg ctaaggtgca gccagctctg ccttcacggt gggaatgtct ctatgtgagc
      901 cctttctggg ctgtgaagaa cgctctgaga agggtcctgg gaccctggat aggccagagc
      961 tgtgctgggc atgtagagac aggagtgggc taaagcagca aaggcactga ccaaggaaga
     1021 gttcagagag gagcgtggaa tatggggagg ggttcatagt aagagagagc aggcagtgga
     1081 gagtaaatag tcactgagcc ggggtttatg gggtttgtag gagcttactc agagaaagta
     1141 gatgagagat gccatgccag tctgagtatc acagagcccc aggctctcct gggaacggaa
     1201 ctgtgagggc cagaaggtca gcaagggagg ttagggagag ttccttttgt actgactcag
     1261 catttatcct gctcccaggg ggcaatgggg gccagtgagg gatgcagagc aaggcagtga
     1321 tgtggcaggc agttcctgtt gtgaaagagc tgggaaggga gcgggctggg cctggtacgt
     1381 acagcaggcc atttctgagg gtccgagtgc tgtctaggag gtgcagtgag acttcagtga
     1441 tcagccagaa cagaagctaa gcggggtggg gactgcgagt tcaggcttct gggtcttgca
     1501 aatatccaga atgctaaatc ctcagaaccc cagggtggcc attttcagag tgggttttgt
     1561 cctttgggca cttgtgcaga ctccaatatc cagagggata aggatggtac tcttcagtac
     1621 ccttagtgag aggacacttt tctctgaagg gcttgaatgt gccgagccat tacctgaagg
     1681 aaggaaatga ctccagggac ataggatggg cccagcacaa ctcacctgct acagagaaag
     1741 gtcccctccc tggtctcctt agagatcctg tttccctggc tgaggaagct agggtggatc
     1801 tttgtgtaag tgggtgtgga tgctaactgg aaaacaaaag gtcacttact gttagacctc
     1861 ggggtaccat ggaagagatg atcactgagt gtgcccttac atggggacca gctgagaatg
     1921 gggctaccac tagctcgaga ccatgataca gggaataagt gtgcatttgg gggtagggag
     1981 tggctcagaa tactcttaac caaagcagag gtttgctccc acaggaaggt gaggtcagaa
     2041 ggccttaggg agctgccagg ggctagggtt ggcaccatct cccaggctgt gtctttaagg
     2101 agatgataat cagagggata gaaccttgca aaagtgggcc agtcttggga atactataga
     2161 ggaatagcct tctggaacat tctgtgtctc ataggacctg cctggggatc cagccccagt
     2221 gccagcacat ataccgactg gggcagtgaa tagatagtac actttgttac atgggctggg
     2281 gggaacatgg cccatgtcct ggaggggact ttatgacaga catccaaaaa tccagtgaga
     2341 gggcttctag atttgtctcc aaaggttata gttctaacat gagcccttag gaaatccagc
     2401 atggttctcc ctgtgtgccc tggtttggtt agagagctct agcggtctcc tgtcccacag
     2461 aataccagcc agcccctgcc ctacgtcgtg cctcgggctg agggtgattc agaggcaggt
     2521 gcctgtgaca gtggatgcaa ttagatctaa tgggacggag gcctttctcg tcgccctcgc
     2581 tccatgccca cccccgcctc cctcaggcac agcaggcgtg gagaggatgc gcaggaggta
     2641 ggaggtgggg gacccagagg ggctttgacg tcagcctggc ctttaaagag ggcgcctgcc
     2701 tggcgagggc tgtggagaca gaactcggga ccaccagagg aattcgatat caagctgcta
     2761 gcgtttaaac gggccctcta gactcgagct caagcttgcc accatggacg tggtgaatca
     2821 gctggtggct gggggtcagt tccgggtggt caaggagccc cttggcttcg tgaaggtgct
     2881 gcagtgggtc tttgccatct tcgcctttgc tacgtgcggc agctacaccg gagagcttcg
     2941 gctgagcgtg gagtgtgcca acaagacgga gagtgccctc aacatcgaag tcgaatttga
     3001 gtacccattc aggctgcacc aagtgtactt tgatgcaccc tcctgcgtta aagggggcac
     3061 taccaagatc ttcctagttg gtgactactc ctcctcggct gaattctttg tcaccgtggc
     3121 tgtgtttgcc ttcctctact ccatgggggc cctggccacc tacatcttcc tgcagaacaa
     3181 gtaccgagag aacaacaaag ggccaatgat ggacttcctg gccacagcag tgttcgcttt
     3241 catgtggcta gttagctcat ccgcctgggc caaaggcctg tccgatgtga agatggccac
     3301 tgacccagag aacattatca aggagatgcc tatgtgccgc cagacagatc tagccaccat
     3361 gggaagtaaa ggagaagaac ttttcactgg agttgtccca attcttgttg aattagatgg
     3421 tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg caacatacgg
     3481 aaaacttacc cttaaattta tttgcactac tggaaaacta cctgttcctt ggccaacact
     3541 tgtcactact ttaacttatg gtgttcaatg cttttcaaga tacccagatc atatgaaacg
     3601 gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaaagaa ctatattttt
     3661 caaagatgac gggaactaca agacacgtgc tgaagtcaag tttgaaggtg atacccttgt
     3721 taatagaatc gagttaaaag gtattgattt taaagaagat ggaaacattc ttggacacaa
     3781 attggaatac aactataacg atcaccaggt gtacatcatg gcagacaaac aaaagaatgg
     3841 aatcaaagct aacttcaaaa ttagacacaa cattgaagat ggaggcgttc aactagcaga
     3901 ccattatcaa caaaatactc caattggcga tgggcccgtc cttttaccag acaaccatta
     3961 cctgtttaca acttctactc tttcgaaaga tcccaacgaa aagagagacc acatggtcct
     4021 tcttgagttt gtaacagctg ctgggattac acatggcatg gatgaactat acaaaaccgg
     4081 tggagctcga atcacatgca aggaactgag ggaccctgtg acttcaggac tcaacacctc
     4141 ggtggtgttt ggcttcctga acctggtgct ctgggttggc aacctatggt tcgtgttcaa
     4201 ggagacaggc tgggccgccc cattcatgcg cgcacctcca ggcgccccag aaaagcaacc
     4261 agctcctggc gatgcctacg gcgatgcggg ctatgggcag ggccccggag gctatgggcc
     4321 ccaggactcc tacgggcctc agggtggtta tcaacccgat tacgggcagc cagccagcgg
     4381 cggtggcggt ggctacgggc ctcagggcga ctatgggcag caaggctacg gccaacaggg
     4441 tgcgcccacc tccttctcca atcagatgtc ggtcgactct agagcctcga ctgtgccttc
     4501 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     4561 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     4621 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     4681 tagcaggcat gctggggatg cggtgggctc tatggcttct gagggtcgag agatctacgg
     4741 gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt
     4801 gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt
     4861 ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac
     4921 ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct
     4981 cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt
     5041 gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg
     5101 tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg
     5161 gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat
     5221 tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc
     5281 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     5341 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     5401 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     5461 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     5521 gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct
     5581 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     5641 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     5701 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     5761 ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt
     5821 ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac
     5881 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     5941 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     6001 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     6061 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     6121 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     6181 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     6241 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     6301 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     6361 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     6421 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     6481 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     6541 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     6601 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     6661 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     6721 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     6781 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     6841 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     6901 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     6961 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     7021 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     7081 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     7141 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     7201 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     7261 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     7321 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     7381 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     7441 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     7501 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     7561 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     7621 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     7681 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     7741 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     7801 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     7861 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     7921 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     7981 ttgctca
//
