LOCUS       pAAV-pChAT-tdTom        6278 bp DNA     circular SYN 01-DEC-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6278)
  AUTHORS   Casey Maguire
  TITLE     Direct Submission
  JOURNAL   Exported Jan 25, 2023 from SnapGene 6.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6278
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     repeat_region   2..131
                     /label=AAV2 ITR
     primer_bind     complement(13..24)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     74..85
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    136..310
                     /label=CMV enhancer (Partial)
     promoter        311..1204
                     /label=human ChAT promoter
     primer_bind     311..327
                     /label=5' ChAT p
     primer_bind     complement(1195..1204)
                     /label=3' ChAT p
     regulatory      1210..1219
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1216..2646
                     /codon_start=1
                     /label=tdTomato
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     misc_feature    2671..3259
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2724..2744)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3142..3153)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3302..3423
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3313..3332)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3367..3386
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3585..3714
                     /label=AAV2 ITR
     primer_bind     complement(3631..3642)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3692..3703
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3832..3849)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4003..4591)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4083..4102)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4762..5622)
                     /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     5385..5404
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5623..5727)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5753..6208)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5890..5911)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6102..6121
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt
       61 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
      121 taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacgt agccatgctc
      181 taggaagagt accattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      241 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      301 aagtgtatca cccgggagca gggggtgggg aaggaggagc agtggctggt ggcaagtctg
      361 agccaggaca tagccgatga cagcctgtga atgaccggcc agactggggt tggaccttcg
      421 gggcagagca ggcagccctt ggaaagccag gctggtagga gcatgggtgg gaggcacaaa
      481 acacagacaa acagacatat atacacatat atcctctagg catctccgag gagtgatgga
      541 ggagagagtt ggtaggtggg accacaggaa accatcccac ccagctctgc cttcttcaag
      601 ctcagggtcc ctgaagcctg actagtgtct cccgaatggg ggaggggggg gctctgggca
      661 gagcaaaagg aggcattggc tgcatgtctt agggccttta ccaactgacc ttgtccttgc
      721 tgtcctctca aaagactgcc atgctccggt gaatcctcac caggacgcca cctctttcac
      781 tttgcagttg ggggcagccc ctacgaaccc acagcactgg gaggcgtggg aacaagcctg
      841 tggagagtgt ccctgcactg tgccagcaag ggctttacat tctctgccca gcactgcaag
      901 gcagccccag gctcctccat ccttgggcct ggttcatcca tctttctgga ctctctgttc
      961 ctggcaacaa ccctgccagt cccgatacat gggcggtgct tgcctctggg taggggtggt
     1021 gttgggtggg gagaggggtg aggaaattag ctaaacctca gctttagact tgagagaata
     1081 gagggtgggc tggtggaaaa catgagtatg tgcctgtgga ggcttaggtt cagggggatc
     1141 tcagagggta gcaattgtga cccacagcct aataataaca gtctttgccc tcttggcctg
     1201 gctcgtaccg ccaccatggt gagtaagggc gaggaagtga tcaaagagtt catgcggttt
     1261 aaggtgagaa tggaaggaag catgaacggc cacgagttcg aaattgaggg agaaggagag
     1321 ggacggccct acgagggcac ccagacagcc aagctgaaag tgacaaaggg cgggcctctg
     1381 ccattcgctt gggacatcct gagcccacag tttatgtacg gctccaaggc ctatgtgaaa
     1441 catccagctg acattcccga ttataagaaa ctgagcttcc ccgaggggtt taagtgggaa
     1501 agagtgatga acttcgagga cggaggcctg gtgactgtga cccaggacag ctccctgcag
     1561 gatgggaccc tgatctacaa ggtgaaaatg agagggacaa attttccccc tgatggacct
     1621 gtgatgcaga agaaaactat gggatgggag gcctccaccg aaaggctgta tccacgcgac
     1681 ggggtgctga aaggagaaat ccaccaggct ctgaagctga aagatggggg acattacctg
     1741 gtggagttca agacaatcta catggccaag aaacctgtgc agctgccagg ctactattac
     1801 gtggacacaa aactggatat cacttcacac aacgaggact acactattgt ggagcagtat
     1861 gaacggagcg aggggagaca ccatctgttc ctgggccatg ggactggaag taccggctca
     1921 gggtctagtg gaaccgcctc aagcgaggat aacaatatgg ctgtgatcaa agagttcatg
     1981 aggtttaagg tgcgcatgga gggcagcatg aatgggcacg aatttgagat tgaaggagag
     2041 ggcgaaggga ggccttacga gggcacacag actgccaagc tgaaagtgac caagggagga
     2101 ccactgcctt tcgcttggga tatcctgtct cctcagttta tgtacggaag taaggcctat
     2161 gtcaagcatc ccgctgacat tcctgattac aagaaactgt ctttcccaga gggctttaag
     2221 tgggagagag tgatgaattt tgaagatgga ggcctggtga ccgtgacaca ggactcctct
     2281 ctgcaggatg gcactctgat ctacaaagtc aaaatgcgcg gcaccaattt tccacccgat
     2341 gggcccgtga tgcagaagaa aacaatgggg tgggaggcca gcactgaacg gctgtatcct
     2401 agagacggag tgctgaaggg cgaaatccac caggccctga agctgaaaga cggcggccac
     2461 tacctggtgg agttcaaaac catctacatg gccaagaaac cagtgcagct gcccggctat
     2521 tactatgtgg acaccaagct ggatatcaca tcccacaatg aagactacac cattgtggaa
     2581 cagtatgaga ggtctgaagg acgccaccat ctgtttctgt acggcatgga tgagctgtat
     2641 aagtaagaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa
     2701 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     2761 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     2821 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     2881 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     2941 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     3001 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     3061 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     3121 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     3181 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     3241 ctttgggccg cctccccgca tcgataccgt cgacccgggc ggccgcttcg agcagacatg
     3301 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     3361 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     3421 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt
     3481 ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatct tcctagagca
     3541 tggctacgta gataagtagc atggcgggtt aatcattaac tacaaggaac ccctagtgat
     3601 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     3661 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcat
     3721 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc attaatgaat
     3781 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3841 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3901 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3961 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4021 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4081 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4141 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4201 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4261 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4321 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4381 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4441 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4501 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4561 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4621 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4681 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4741 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4801 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4861 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4921 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4981 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5041 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5101 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5161 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5221 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5281 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5341 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5401 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5461 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5521 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5581 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5641 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5701 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta
     5761 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     5821 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     5881 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     5941 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     6001 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     6061 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     6121 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     6181 gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg
     6241 gaagggcgat cggtgcgggc ctcttcgcta ttacgcca
//
