LOCUS       sequence-484983-        6152 bp    DNA     circular SYN 28-AUG-2025
DEFINITION  Expression of GFP in cortical FOXP2_RORB neurons in marmoset.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6152)
  TITLE     Marmoset Enhancers
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6152)
  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            "Unpublished"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-2xcHS4-cjFOXP2_RORB-E4-CMVmini-EGFP-miR-WPRE-hGHpA
FEATURES             Location/Qualifiers
     source          1..6152
                     /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     2165..2189
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2287..2296
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2293..2295,2296..2298,2299..3012)
                     /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(2293..2295,2296..2298,2299..3009)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2338..2359)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2599..2618)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2946..2967
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3226..3814
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3279..3299)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3697..3708)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     repeat_region   4065..4205
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4065..4194
                     /label=AAV2 ITR
     rep_origin      4280..4735
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4367..4386)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4577..4598
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4752..4771)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4871..4893
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4931..4949)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5017..5121
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5122..5190,5191..5982)
                     /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(5340..5359)
                     /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 gcgtacaggg taattgcgcg ccgagctcac ggggacagcc cccccccaaa
      841 gcccccaggg atgtaattac gtccctcccc cgctaggggg cagcagcgag ccgcccgggg
      901 ctccgctccg gtccggcgct ccccccgcat ccccgagccg gcagcgtgcg gggacagccc
      961 gggcacgggg aaggtggcac gggatcgctt tcctctgaac gcttctcgct gctctttgag
     1021 cctgcagaca cctgggggat acggggaaaa agcttgagct cacggggaca gccccccccc
     1081 aaagccccca gggatgtaat tacgtccctc ccccgctagg gggcagcagc gagccgcccg
     1141 gggctccgct ccggtccggc gctccccccg catccccgag ccggcagcgt gcggggacag
     1201 cccgggcacg gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt
     1261 gagcctgcag acacctgggg gatacgggga aaaagcttat cgacggccga cgccgtttaa
     1321 ttaagtgtct agactgcaga gggcccgaat tcgcatgcaa agcatttttg gaaaggcctc
     1381 ctagaggaag taggacttga aggaagacag atcagtggtt ttcaagtata actccacctt
     1441 aaaacaatct ggagagcttt actaaaacac acacacacac gcatgtacgc acactcacac
     1501 cccttccccg tgcctgggtc ccactctgat cctattgaat cataatctct ggggacaggc
     1561 tctgaaactg catttttcaa aacttgacag gtgagtctaa tgtgcaacca ggattgtgaa
     1621 tcgcatagtc tcaaaaagca gaaatagaag gaggagaatt ctaagttgaa gaaacgtcat
     1681 aaatgacggc actgggaagg aaagaagagg cacgacttcg ggatgttgac taatgcagca
     1741 tgatcacagc attgaaaacc catggagatg tgggacagat ggggctagaa acataggcag
     1801 ggactttgga gggccctgag gctgttgggg cctcagtgta ggttggtgct tgatttccac
     1861 ctactcaccc tttttacttt caagtttctg ggttgaagag ccagatgggg caagggacca
     1921 cttttagata ggactttctg aaaacagata gaaaagagtg aagcctatat cttgaggcca
     1981 catgacccag attttcctcc atctgtttgt aggagatgaa gccggagcag aaaagcagaa
     2041 atctcccctt gggttccggt atctgttgcc aactgatcag tggggtggct tcactctctg
     2101 gttatggttt actagtgata tcgtataagc tttaggcgtg tacggtgggc gcctataaaa
     2161 gcagagctcg tttagtgaac cgtcagatcg cctggagcaa ttccacaaca cttttgtctt
     2221 ataccaactt tccgtaccac ttcctaccct cgtaaaggat ccttcgaagc tagcagcgct
     2281 gttaacgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     2341 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     2401 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     2461 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     2521 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     2581 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     2641 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     2701 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     2761 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     2821 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     2881 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     2941 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     3001 ctgtacaagt aagtcgacgg cgcgccaccg gtcaaacacc attgtcacac tccatactat
     3061 caaacaccat tgtcacactc cagatcaaca aacaccattg tcacactcca cgccggcggg
     3121 taccagtgaa ttctaccagt gccatagatc aaagtgaatt ctaccagtgc catatactat
     3181 agtgaattct accagtgcca tacaagctta tcgatctcga tcgataatca acctctggat
     3241 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     3301 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     3361 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     3421 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     3481 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     3541 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     3601 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc
     3661 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     3721 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     3781 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat aagatctacg ggtggcatcc
     3841 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag
     3901 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat
     3961 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc
     4021 ctgcggggtc tattgggaac cacacgtgcg gaccgagcgg ccgcaggaac ccctagtgat
     4081 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     4141 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     4201 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     4261 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4321 gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc
     4381 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4441 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     4501 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4561 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg
     4621 ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag
     4681 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     4741 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     4801 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4861 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     4921 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     4981 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5041 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5101 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5161 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5221 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5281 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5341 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5401 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5461 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5521 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5581 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5641 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5701 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5761 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5821 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5881 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5941 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6001 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6061 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6121 tcagaccccg tagaaaagat caaaggatct tc
//
