LOCUS       sequence-484986-        5450 bp    DNA     circular SYN 28-AUG-2025
DEFINITION  Expression of GFP in striatal D2 medium spiny neurons in marmoset.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5450)
  TITLE     Marmoset Enhancers
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5450)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 28, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-cjD2-E8-CMVmini-EGFP-miR-WPRE3-hGHpA
FEATURES             Location/Qualifiers
     source          1..5450
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   651..780
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     primer_bind     1809..1833
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1931..1940
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1937..1939,1940..1942,1943..2656)
                     /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             join(1937..1939,1940..1942,1943..2653)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1982..2003)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2243..2262)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2590..2611
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2912..2932)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   3363..3503
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3363..3492
                     /label=AAV2 ITR
     rep_origin      3578..4033
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3665..3684)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3875..3896
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4050..4069)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4169..4191
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4229..4247)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4315..4419
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4420..4488,4489..5280)
                     /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(4638..4657)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
      661 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
      721 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      781 gcggccgcac gcgtttaatt aagtgtctag actgcagagg gcccgaattc gcatgctaca
      841 aatgaagcgt aaatctaccc agtgagggag gaatattctt accaccttga gaataaccgc
      901 ggtgatgaca aacaaggggc tggcacccac gggctcaggc tctggggagc tgtcagggcg
      961 taatttgcat gctaaataca gtatttttag caccaaagtt tggagcactt atcttgccct
     1021 gaacagttaa ttatggactt ttgtcttctc ccgaacctaa aggtagtgct gagccctggg
     1081 agaggctcct ctgtccaaga gcaccctgat tctggaaggt gggcaaaaca ggcccctagt
     1141 ctaacttagg gggaccactg gcccagtgac taccttctgg ggaaggcatc aggtccccaa
     1201 gagggataga agccccagga acccaaccca tctccattgc acacatacca tgctcaaagt
     1261 ctgtgaaaga tgttgacctg ggtggacgct taaaagcata acccacgatt aggaatctta
     1321 cgagggtata caggcttatc agctgtgagg attggaagag aaaacaaagg gaaacagagg
     1381 aaagaagaag gaagggaggg agagagagag ggagtgaagc aagacccaga gcagagtgag
     1441 aagagcattg acagggagca gaggggaaga gggcagggca ggggcgggag gcggggcagg
     1501 ggaaagttgc ccacagttgt cacgaggctt cacgtctttc taccagacag cagattgaca
     1561 gctagagtgg gcagggggag ggttgggatc caccctctcc cccctcagca cttcaggggc
     1621 aaagaaatgg gaggagtagg acctgacacg acacgggaac acaaggtgga aggggtggcc
     1681 caggctctga ctctctccag agaggccctg acgatggtct cttgctcttg acggcagggc
     1741 agaaaggaag cctcactagt gatatcgtat aagctttagg cgtgtacggt gggcgcctat
     1801 aaaagcagag ctcgtttagt gaaccgtcag atcgcctgga gcaattccac aacacttttg
     1861 tcttatacca actttccgta ccacttccta ccctcgtaaa ggatccttcg aagctagcag
     1921 cgctgttaac gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     1981 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     2041 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     2101 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     2161 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     2221 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     2281 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     2341 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     2401 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     2461 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     2521 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     2581 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     2641 cgagctgtac aagtaagtcg acggcgcgca ccggtcaaac accattgtca cactccatac
     2701 tatcaaacac cattgtcaca ctccagatca acaaacacca ttgtcacact ccacgccggc
     2761 gggtaccagt gaattctacc agtgccatag atcaaagtga attctaccag tgccatatac
     2821 tatagtgaat tctaccagtg ccatacaagc ttatcgataa tcaacctctg gattacaaaa
     2881 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     2941 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     3001 tgtataaatc ctggttagtt cttgccacgg cggaactcat cgccgcctgc cttgcccgct
     3061 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttagcg ctgctcgaga
     3121 gatctacggg tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc
     3181 cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta
     3241 ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg
     3301 gaagacaacc tgtagggcct gcggggtcta ttgggaacca cacgtgcgga ccgagcggcc
     3361 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     3421 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     3481 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     3541 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     3601 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
     3661 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3721 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3781 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3841 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3901 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc
     3961 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     4021 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     4081 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     4141 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     4201 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     4261 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4321 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4381 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4441 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4501 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4561 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4621 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4681 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4741 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4801 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4861 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     4921 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     4981 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5041 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5101 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5161 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5221 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5281 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5341 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5401 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
//
