LOCUS       sequence-469484-        7370 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent Reporter for Dopaminergic Neuron Differentiation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7370)
  AUTHORS   Oh MS, Hong SJ, Huh Y, Kim KS
  TITLE     Expression of transgenes in midbrain dopamine neurons using the 
            tyrosine hydroxylase promoter.
  JOURNAL   Gene Ther. 2009 Mar;16(3):437-40. doi: 10.1038/gt.2008.148. Epub 
            2008 Sep 18.
  PUBMED    18800154
REFERENCE   2  (bases 1 to 7370)
  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"; doi: 
            "10.1038/gt.2008.148"; journalName: "Gene Ther"; date: "2009-03"; 
            volume: "16"; issue: "3"; pages: "437-40"
COMMENT     Sequence Label: pAAV2.5-THP-GFP/WGA
FEATURES             Location/Qualifiers
     source          1..7370
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      479..1067
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     968..987
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1129..1258
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3856..3865
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3862..3864,3865..3867,3868..4578)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3862..3864,3865..3867,3868..4578)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3907..3928)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4168..4187)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4515..4536
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5245..5721
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5444..5463)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5761..5901
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5761..5890
                     /label=AAV2 ITR
     rep_origin      5976..6431
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6063..6082)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6273..6294
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6448..6467)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6567..6589
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6627..6645)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6713..6817
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6818..6886,6887..7370,1..308)
                     /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(7036..7055)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
       61 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
      121 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
      181 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
      241 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
      301 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
      361 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
      421 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
      481 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
      541 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
      601 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
      661 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
      721 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
      781 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
      841 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
      901 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
      961 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     1021 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     1081 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtcc tgcaggcagc
     1141 tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg cgacctttgg tcgcccggcc
     1201 tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc
     1261 ggccgcacgc gaggcctctt gggattcttc tcatcaaaaa gggcatggga cacagctaaa
     1321 gcgtccaggg ctcctctgtg cccacagatg ccttagatct tggcacaatg tagtcagcca
     1381 gctccgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg
     1441 tgtgtgtgtg tttgcatgta tctcacagac agtgcacaat ggcctggatg tgaacagagg
     1501 caagagtctg ggccagcagt tgtctcccag gagggtccaa agacatcgta ttttcaagtt
     1561 taggccaggt gctcacttgg gtgagctcag acacagacaa aggtctggag agcacacatt
     1621 ccccaccccc acccagcttc tatgcaagca cctccagctg agacaagaaa acgaattaaa
     1681 aagcaatatt tgtgtcagcg taagacattt gccgaaaggt taaatccaca ctcgtggtgc
     1741 tgcacagcag ccccctgtgc aggatttgtt aggcacagct ccctcctacc ccgtgccacc
     1801 tgagcaaatg ccaggctggg tgggctggaa ccagcctggg cttgcctcac cctggaatcc
     1861 ccagcaccct ccaaaggagg accctgggag tgggcataga cgcccttcag gtgtgggcaa
     1921 cagcccccag tcctcaggat gaaaggctaa ggtgcagcca gctctgcctt cacggtggga
     1981 atgtctctat gtgagccctt tctgggctgt gaagaacgct ctgagaaggg tcctgggacc
     2041 ctggataggc cagagctgtg ctgggcatgt agagacagga gtgggctaaa gcagcaaagg
     2101 cactgaccaa ggaagagttc agagaggagc gtggaatatg gggaggggtt catagtaaga
     2161 gagagcaggc agtggagagt aaatagtcac tgagccgggg tttatggggt ttgtaggagc
     2221 ttactcagag aaagtagatg agagatgcca tgccagtctg agtatcacag agccccaggc
     2281 tctcctggga acggaactgt gagggccaga aggtcagcaa gggaggttag ggagagttcc
     2341 ttttgtactg actcagcatt tatcctgctc ccagggggca atgggggcca gtgagggatg
     2401 cagagcaagg cagtgatgtg gcaggcagtt cctgttgtga aagagctggg aagggagcgg
     2461 gctgggcctg gtacgtacag caggccattt ctgagggtcc gagtgctgtc taggaggtgc
     2521 agtgagactt cagtgatcag ccagaacaga agctaagcgg ggtggggact gcgagttcag
     2581 gcttctgggt cttgcaaata tccagaatgc taaatcctca gaaccccagg gtggccattt
     2641 tcagagtggg ttttgtcctt tgggcacttg tgcagactcc aatatccaga gggataagga
     2701 tggtactctt cagtaccctt agtgagagga cacttttctc tgaagggctt gaatgtgccg
     2761 agccattacc tgaaggaagg aaatgactcc agggacatag gatgggccca gcacaactca
     2821 cctgctacag agaaaggtcc cctccctggt ctccttagag atcctgtttc cctggctgag
     2881 gaagctaggg tggatctttg tgtaagtggg tgtggatgct aactggaaaa caaaaggtca
     2941 cttactgtta gacctcgggg taccatggaa gagatgatca ctgagtgtgc ccttacatgg
     3001 ggaccagctg agaatggggc taccactagc tcgagaccat gatacaggga ataagtgtgc
     3061 atttgggggt agggagtggc tcagaatact cttaaccaaa gcagaggttt gctcccacag
     3121 gaaggtgagg tcagaaggcc ttagggagct gccaggggct agggttggca ccatctccca
     3181 ggctgtgtct ttaaggagat gataatcaga gggatagaac cttgcaaaag tgggccagtc
     3241 ttgggaatac tatagaggaa tagccttctg gaacattctg tgtctcatag gacctgcctg
     3301 gggatccagc cccagtgcca gcacatatac cgactggggc agtgaataga tagtacactt
     3361 tgttacatgg gctgggggga acatggccca tgtcctggag gggactttat gacagacatc
     3421 caaaaatcca gtgagagggc ttctagattt gtctccaaag gttatagttc taacatgagc
     3481 ccttaggaaa tccagcatgg ttctccctgt gtgccctggt ttggttagag agctctagcg
     3541 gtctcctgtc ccacagaata ccagccagcc cctgccctac gtcgtgcctc gggctgaggg
     3601 tgattcagag gcaggtgcct gtgacagtgg atgcaattag atctaatggg acggaggcct
     3661 ttctcgtcgc cctcgctcca tgcccacccc cgcctccctc aggcacagca ggcgtggaga
     3721 ggatgcgcag gaggtaggag gtgggggacc cagaggggct ttgacgtcag cctggccttt
     3781 aaagagggcg cctgcctggc gagggctgtg gagacagaac tcgggaccac cagaggaatt
     3841 ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
     3901 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     3961 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     4021 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     4081 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     4141 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     4201 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     4261 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     4321 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     4381 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     4441 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     4501 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     4561 atggacgagc tgtacaagtc cggccggact cagatctcga tgagaaagat gatgagcacc
     4621 atggccctta cgctcggcgc ggctgtcttc ctcgccttcg ccgcggcgac cgcgcaggcc
     4681 cagaggtgcg gcgagcaggg cagcaacatg gagtgcccca acaacctctg ctgcagccag
     4741 tacgggtact gcggcatggg cggcgactac tgcggcaagg gctgccagaa cggcgcctgc
     4801 tggaccagca agcgctgcgg cagccaggcc ggcggggcga cgtgtcccaa caaccactgc
     4861 tgcagccagt acgggcactg cggcttcgga gccgagtact gcggcgccgg ctgccagggc
     4921 ggcccctgcc gcgccgacat caagtgcggc agccagtccg gcggcaagct atgcccgaac
     4981 aacctctgct gcagccagtg gggattctgc ggcctaggtt ccgagttctg cggcggtggc
     5041 tgccagagcg gtgcttgcag caccgacaag ccgtgcggca aggacgccgg cggcagggtt
     5101 tgcactaaca actactgttg tagcaagtgg ggatcctgtg gcatcggccc gggctattgc
     5161 ggtgcaggcc gccagagcgg cggctgtgac gcttaagtcg acctgcagaa gcttgcctcg
     5221 agcagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc agtgcctctc
     5281 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc
     5341 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg
     5401 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc
     5461 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt
     5521 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa
     5581 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc
     5641 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc
     5701 actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc gagcggccgc
     5761 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     5821 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     5881 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     5941 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     6001 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     6061 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     6121 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     6181 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     6241 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     6301 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
     6361 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     6421 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     6481 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     6541 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     6601 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     6661 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     6721 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     6781 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     6841 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     6901 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     6961 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     7021 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     7081 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     7141 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     7201 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     7261 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     7321 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
//
