LOCUS       sequence-411050-        6077 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Direct-expressing iAChSnFR-Venus in astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6077)
  TITLE     Allen Institute Plasmids Aug 2023
REFERENCE   2  (bases 1 to 6077)
  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"
COMMENT     Sequence Label: AiP14534 -
            pAAV-AiE0390m_3xC2-minBglobin-iAChS...
FEATURES             Location/Qualifiers
     source          1..6077
                     /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      988..997
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     994..1053
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     1506..1527
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1630..1651)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2638..2667
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2671..2817
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     primer_bind     complement(2904..2924)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3111..3222
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3340..3480
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3340..3469
                     /label=AAV2 ITR
     rep_origin      3555..4010
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3642..3661)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3852..3873
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4027..4046)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4146..4168
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4206..4224)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4292..4396
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4397..4465,4466..5257)
                     /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(4615..4634)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5428..6016
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5917..5936
                     /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 gcgtgaaagg aaagagcttg ctttcaacct caaaagctag
      181 gaggaaaggg ctctgaaatt tgctcagaat tcccaattca ccattagcct gtttcttcct
      241 ttagcctcaa ggcattctcc gctttttgaa aagatgttaa gaaattcagt cacaatagag
      301 agcctagttt tgaacatgtt tcactcggtc cattgaggtc tgggctccag cctttgtgtg
      361 gggtgaattg agctgagcgg ctagctggtt ggaggaaagg aaagagcttg ctttcaacct
      421 caaaagctag gaggaaaggg ctctgaaatt tgctcagaat tcccaattca ccattagcct
      481 gtttcttcct ttagcctcaa ggcattctcc gctttttgaa aagatgttaa gaaattcagt
      541 cacaatagag agcctagttt tgaacatgtt tcactcggtc cattgaggtc tgggctccag
      601 cctttgtgtg gggtgaattg agctgagcgg ctagctggtt ggaggaaagg aaagagcttg
      661 ctttcaacct caaaagctag gaggaaaggg ctctgaaatt tgctcagaat tcccaattca
      721 ccattagcct gtttcttcct ttagcctcaa ggcattctcc gctttttgaa aagatgttaa
      781 gaaattcagt cacaatagag agcctagttt tgaacatgtt tcactcggtc cattgaggtc
      841 tgggctccag cctttgtgtg gggtgaattg agctgagcgg ctagctggtt ggaggagctc
      901 gggctgggca taaaagtcag ggcagagcca tctattgctt acatttgctt ctgggatcca
      961 gatctttcga agctagcgct accggtcgcc accatggaga cagacacact cctgctatgg
     1021 gtactgctgc tctgggttcc aggttccact ggtgacagat ctgcgaacga caccgtagtt
     1081 gtgggctcga tcatctttac agaagggatt atcgtcgcaa acatggtggc agagatgatt
     1141 gaggcgcata cagaccttaa ggtggttcgc aaactgaacc ttggcggggt caacgttaac
     1201 tttgaagcca ttaaacgcgg aggtgcgaat aatggtattg acatttacgt ggagtacact
     1261 gggcacggtc ttgtggatat tctggggttc ccggagccga acgtctatat caccgccgac
     1321 aagcagaaga acggcatcaa ggcgaacttc aagatccgcc acaacgtgga ggacggcagc
     1381 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1441 cccgacaacc actacctgag ctatcagtcc gtgctgagca aagaccccaa cgagaagcgc
     1501 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catgaacgag
     1561 ctgtacaagg gcggtaccgg agggagcatg agcaagggcg aggagctgtt caccggggtg
     1621 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgcgcggc
     1681 gagggcgagg gcgatgccac caacggcaag ctgaccctga agctcctgtg caccaccggc
     1741 aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcgt gcagtgcttc
     1801 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     1861 tacgtccagg agcgcaccat cagcttcaag gacgacggca cctacaagac ccgcgccgag
     1921 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1981 gaggacggca acatcctggg gcacaagctg gagtacaact ttccgccgcc cgcgactact
     2041 gatccagaag gtgcatacga aaccgtgaag aaggagtaca aacgtaaatg gaatattgta
     2101 tggctcaaac cactgggatt caacaatacg tatacgctta ccgttaaaga cgaactggcg
     2161 aaacagtata accttaaaac cttcagtgac ttagcgaaaa tctcggataa gctgattctg
     2221 ggtgcaacga tgttcttttt agaagggccc gatggttacc caggcctgca aaaactgtac
     2281 aatttcaaat tcaagcacac caaaagcatg gacatgggta ttcgctatac cgccattgat
     2341 aataacgaag ttcaggtaat tgatgcctgg gccactgatg gcttgctggt gagccacaaa
     2401 ttaaaaattc tggaggatga taaagcgttc ttcccgccgt attatgctgc ccccatcatc
     2461 cgtcaggatg tcttagataa gcatcctgaa ctgaaggacg tgctgaacaa actcgcgaat
     2521 caaatttcac tggaagaaat gcagaaactg aattacaagg tggacggtga gggtcaggac
     2581 ccagcgaaag tagctaagga gtttttgaaa gagaaaggtt taattctgca ggtcgacgaa
     2641 caaaaactca tctcagaaga ggatctgaat gctgtgggcc aggacacgca ggaggtcatc
     2701 gtggtgccac actccttgcc ctttaaggtg gtggtgatct cagccatcct ggccctggtg
     2761 gtgctcacca tcatctccct tatcatcctc atcatgcttt ggcagaagaa gccacgttag
     2821 ggcgcgccgc ggccgcgaat tcgatatcat aatcaacctc tggattacaa aatttgtgaa
     2881 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     2941 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     3001 tcctggttag ttcttgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     3061 ggggctcggc tgttgggcac tgacaattcc gtggctcgag agatcttcga ctgtgccttc
     3121 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3181 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3241 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3301 tagcaggcat gagatctcac gtgcggaccg agcggccgca ggaaccccta gtgatggagt
     3361 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     3421 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     3481 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     3541 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3601 gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc
     3661 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3721 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     3781 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3841 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta
     3901 ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat
     3961 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     4021 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     4081 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     4141 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     4201 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     4261 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4321 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4381 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4441 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4501 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4561 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4621 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4681 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     4741 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4801 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4861 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4921 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     4981 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5041 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5101 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5161 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5221 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5281 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5341 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5401 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5461 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5521 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5581 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5641 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5701 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5761 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5821 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5881 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5941 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6001 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6061 gccttttgct cacatgt
//
