LOCUS       sequence-443644-        5515 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent expression of SYFP2 under the DLX2.0 enhancer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5515)
  TITLE     Striosomes Target Nigral Dopamine-Containing Neurons via Direct-D1 
            and Indirect-D2 Pathways Paralleling Classic Direct-Indirect Basal 
            Ganglia Systems
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5515)
  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            "Unpublished"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: AiP14533-pAAV-DLX2.0-minBG-DIO-SYFP2-WPRE-HGHp...
FEATURES             Location/Qualifiers
     source          1..5515
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     protein_bind    complement(626..659)
                     /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)."
     protein_bind    complement(710..743)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(761..1471,1472..1474,1475..1477))
                     /codon_start=1
                     /product="monomeric optimized variant of EYFP, identical to
                     mVenus but with L68V (Kremers et al., 2006)"
                     /label=SYFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(803..824)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1152..1171
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1411..1432
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1474..1483
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1500..1533
                     /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)."
     protein_bind    1584..1617
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1642..2230
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1695..1715)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2113..2124)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2262..2738
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2461..2480)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2778..2918
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2778..2907
                     /label=AAV2 ITR
     rep_origin      2993..3448
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3080..3099)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3290..3311
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3465..3484)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3584..3606
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3644..3662)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3730..3834
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3835..3903,3904..4695)
                     /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(4053..4072)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4866..5454
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5355..5374
                     /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 gcgtggtacc ctaaataaag atggcttttt agtattaaaa
      181 gtggaagaaa attacaggta attatctttg acggtaaaaa cgctgtaatc agcgggctac
      241 atgaaaaatt actctaatta tggctgcatt taagagaatg gctaaataaa gatggctttt
      301 tagtattaaa agtggaagaa aattacaggt aattatcttt gacggtaaaa acgctgtaat
      361 cagcgggcta catgaaaaat tactctaatt atggctgcat ttaagagaat ggctaaataa
      421 agatggcttt ttagtattaa aagtggaaga aaattacagg taattatctt tgacggtaaa
      481 aacgctgtaa tcagcgggct acatgaaaaa ttactctaat tatggctgca tttaagagaa
      541 tggagctcgg gctgggcata aaagtcaggg cagagccatc tattgcttac atttgcttct
      601 gggatcctct agagtcgact ccggaataac ttcgtatagg atactttata cgaagttatg
      661 cagaatggta gctggattgt agctgctatt agcaatatga aacctcttaa taacttcgta
      721 tagcatacat tatacgaagt tatggcgcgc cgtcgactta cttgtacagc tcgtccatgc
      781 cgagagtgat cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt
      841 tggggtcttt gctcagcttg gactggtagc tcaggtagtg gttgtcgggc agcagcacgg
      901 ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc gagctgcacg ccgccgtcct
      961 cgatgttgtg gcggatcttg aagttggcct tgatgccgtt cttctgcttg tcggcggtga
     1021 tatagacgtt gtggctgttg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct
     1081 ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca
     1141 cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt
     1201 agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggg
     1261 cgaagcactg cacgccgtag cccagggtgg tcacgagggt gggccagggc acgggcagct
     1321 tgccggtggt gcagatcagc ttcagggtca gcttgccgta ggtggcatcg ccctcgccct
     1381 cgccggacac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca
     1441 ccaccccggt gaacagctcc tcgcccttgc tcaccatggt ggcgaccggt agcgctagca
     1501 taacttcgta taaagtatcc tatacgaagt tatttgcctt aacccagaaa ttatcactgt
     1561 tattctttag aatggtgcaa agaataactt cgtataatgt atgctatacg aagttatgaa
     1621 ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     1681 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     1741 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     1801 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     1861 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     1921 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     1981 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     2041 tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc
     2101 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     2161 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     2221 gcctccccgc atcgataccg agcgctgctc gagagatcta cgggtggcat ccctgtgacc
     2281 cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc
     2341 taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt
     2401 ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg
     2461 tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct
     2521 cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca
     2581 tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca
     2641 ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg
     2701 gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag gtaaccacgt
     2761 gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     2821 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     2881 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     2941 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     3001 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3061 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3121 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3181 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     3241 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3301 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
     3361 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3421 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     3481 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     3541 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     3601 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     3661 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     3721 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3781 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     3841 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     3901 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     3961 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     4021 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     4081 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     4141 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4201 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     4261 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4321 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     4381 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     4441 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     4501 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     4561 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4621 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     4681 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     4741 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4801 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4861 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4921 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4981 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5041 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5101 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5161 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5221 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5281 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5341 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5401 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5461 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
