LOCUS       sequence-446776-        5392 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AiE1421m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in striatal Pvalb+/Pthlh+ neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5392)
  TITLE     Enhancer AAV toolbox for accessing and perturbing striatal cell 
            types and circuits
REFERENCE   2  (bases 1 to 5392)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AiP15045 -
            pAAV-AiE1421m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5392
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      745..754
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(751..753,754..756,757..1467)
                     /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(1057..1076)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1404..1425
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1477..1533
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1546..1611
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1642..2019
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2106..2126)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2313..2424
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2542..2682
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2542..2671
                     /label=AAV2 ITR
     misc_feature    2689..2716
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    2717..2744
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      2813..3268
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2900..2919)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3110..3131
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3285..3304)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3404..3426
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3464..3482)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3550..3654
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3655..3723,3724..4515)
                     /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(3873..3892)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4686..5274
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5175..5194
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5331..5358
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    5359..5386
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgaatgt ggtgggatga atgctgtcta gcacagtctt
      181 ctggtctcca ggtctccaca gcacagcctg tacttattta caataacctg tatctgttct
      241 cagactggtg aattcagagt actatttatc tcagtgccca agaaccaaag agattctcaa
      301 gagccaactc acagttgcac cttttaaatg ccgtgtgaac tagaattcaa gctcaccaac
      361 acccagaggc aaagcatctg cagcctagtt tccatcggaa gctgggagca ctttgttcat
      421 cacagctcag aatagataca ttaattaatt ggcaaagatg gagccataag agagaccagc
      481 tgccttgtct ggcctcagtg tgagaggatg tgtctagtct cccagagact tgatgtgcca
      541 gggtggagga atacggtagc ggagttgggg tggggggaac tcagagtaga aggggagggg
      601 ctgggatgaa ggattatggg aggggctgac agggagaggg tcagtgagtg ggagctcggg
      661 ctgggcataa aagtcagggc agagccatct attgcttaca tttgcttctg ggatccagat
      721 ctttcgaagc tagcgctacc ggtcgccacc atggtctcca agggcgagga gctgttcacc
      781 ggggtggtgc ccatcctggt cgagctggac ggcgatgtca acggccacaa gttcagcgtg
      841 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc
      901 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcgtgcag
      961 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1021 gaaggctacg tccaggagcg caccatcttc ttcaaagacg acggcaacta caagacccgc
     1081 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1141 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     1201 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac
     1261 aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     1321 gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa gctgagcaaa
     1381 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     1441 actctcggca tggacgagct gtacaagggc tctggtgcta ccaacttctc actgttgaaa
     1501 caggcagggg atgtagagga gaatccaggg cctggtgcta gtggagacta caaagaccat
     1561 gacggagatt ataaagatca tgacatcgat tacaaggatg acgatgacaa gtccggactc
     1621 agatctggag gctccggagg cccagagcca gcgaagtctg ctcccgcccc gaaaaagggc
     1681 tccaagaagg cggtgactaa ggcgcagaag aaaggcggca agaagcgcaa gcgcagccgc
     1741 aaggagagct attccatcta tgtgtataag gttctgaagc aggtccaccc tgacaccggc
     1801 atttcgtcca aggccatggg catcatgaac tcgtttgtga acgacatttt cgagcgcatc
     1861 gcaggtgagg cttcccgcct ggcgcattac aacaagcgct cgaccatcac ctccagggag
     1921 atccagacgg ccgtgcgcct gctgctgcct ggggagttgg ccaagcacgc cgtgtccgag
     1981 ggtactaagg ccatcaccaa gtacaccagc gctaagtaat gaggcgcgcc gcggccgcga
     2041 attcgatatc ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     2101 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     2161 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt agttcttgcc
     2221 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     2281 actgacaatt ccgtggctcg agagatcttc gactgtgcct tctagttgcc agccatctgt
     2341 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2401 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2461 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgagatctc
     2521 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     2581 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     2641 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgccta tcagcacaca
     2701 attgcccatt atacgcgcgt ataatggact attgtgtgct gatatgatgc ggtattttct
     2761 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     2821 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2881 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2941 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3001 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     3061 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3121 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
     3181 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3241 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     3301 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     3361 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     3421 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     3481 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     3541 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3601 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     3661 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     3721 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     3781 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     3841 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     3901 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     3961 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4021 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     4081 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4141 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     4201 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     4261 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     4321 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     4381 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4441 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     4501 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     4561 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4621 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4681 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4741 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4801 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     4861 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4921 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4981 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5041 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5101 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5161 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5221 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5281 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca tatcagcaca
     5341 caattgccca ttatacgcgc gtataatgga ctattgtgtg ctgataacat gt
//
