LOCUS       Exported                6443 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses iAChSnFR under CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV.CAG.iAChSnFR
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6443)
  TITLE     A genetically encoded fluorescent sensor for in vivo acetylcholine 
            detection in mice, fish, worms and flies. (iAChSnFR)
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6443)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6443
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     enhancer        235..538
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        540..817
                     /label=chicken beta-actin promoter
     primer_bind     1074..1093
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     sig_peptide     1148..1210
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
     primer_bind     1660..1681
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1784..1805)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2792..2821
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2825..2971
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     polyA_signal    2976..3110
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3000..3019)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3054..3073
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3196..3213)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3202..3409
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3439..3579
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3439..3568
                     /label=AAV2 ITR
     rep_origin      3675..4130
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3762..3781)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3972..3993
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4197..4216)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4316..4338
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4376..4394)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4462..4566
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4567..5427
                     /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(4785..4804)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5598..6186
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6087..6106
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6340..6357
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtagatc tgaattcggt accctagtta
      181 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      241 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      301 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      361 ggactattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      421 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      481 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      541 cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc cacccccaat
      601 tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg gggggggggg
      661 gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc ggagaggtgc
      721 ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg
      781 gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcga cgctgccttc
      841 gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt
      901 tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg
      961 tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagc
     1021 tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt
     1081 gctggttatt gtgctgtctc atcattttgg caaagaattc ctcgaagatc cgaaggggtt
     1141 caagcttatg gagacagaca cactcctgct atgggtactg ctgctctggg ttccaggttc
     1201 cactggtgac agatctgcga acgacaccgt agttgtgggc tcgatcatct ttacagaagg
     1261 gattatcgtc gcaaacatgg tggcagagat gattgaggcg catacagacc ttaaggtggt
     1321 tcgcaaactg aaccttggcg gggtcaacgt taactttgaa gccattaaac gcggaggtgc
     1381 gaataatggt attgacattt acgtggagta cactgggcac ggtcttgtgg atattctggg
     1441 gttcccggag ccgaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggcgaa
     1501 cttcaagatc cgccacaacg tggaggacgg cagcgtgcag ctcgccgacc actaccagca
     1561 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     1621 gtccgtgctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     1681 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggcggta ccggagggag
     1741 catgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1801 cgacgtaaac ggccacaagt tcagcgtgcg cggcgagggc gagggcgatg ccaccaacgg
     1861 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1921 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     1981 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcagctt
     2041 caaggacgac ggcacctaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     2101 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     2161 gctggagtac aactttccgc cgcccgcgac tactgatcca gaaggtgcat acgaaaccgt
     2221 gaagaaggag tacaaacgta aatggaatat tgtatggctc aaaccactgg gattcaacaa
     2281 tacgtatacg cttaccgtta aagacgaact ggcgaaacag tataacctta aaaccttcag
     2341 tgacttagcg aaaatctcgg ataagctgat tctgggtgca acgatgttct ttttagaagg
     2401 gcccgatggt tacccaggcc tgcaaaaact gtacaatttc aaattcaagc acaccaaaag
     2461 catggacatg ggtattcgct ataccgccat tgataataac gaagttcagg taattgatgc
     2521 ctgggccact gatggcttgc tggtgagcca caaattaaaa attctggagg atgataaagc
     2581 gttcttcccg ccgtattatg ctgcccccat catccgtcag gatgtcttag ataagcatcc
     2641 tgaactgaag gacgtgctga acaaactcgc gaatcaaatt tcactggaag aaatgcagaa
     2701 actgaattac aaggtggacg gtgagggtca ggacccagcg aaagtagcta aggagttttt
     2761 gaaagagaaa ggtttaattc tgcaggtcga cgaacaaaaa ctcatctcag aagaggatct
     2821 gaatgctgtg ggccaggaca cgcaggaggt catcgtggtg ccacactcct tgccctttaa
     2881 ggtggtggtg atctcagcca tcctggccct ggtggtgctc accatcatct cccttatcat
     2941 cctcatcatg ctttggcaga agaagccacg ttagggatcc agacatgata agatacattg
     3001 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     3061 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     3121 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt tagtcgacta
     3181 gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     3241 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     3301 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     3361 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggag agatctcacg
     3421 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     3481 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     3541 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg ctgcagccag
     3601 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gtagcctgaa
     3661 tggcgaatgg cgcgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3721 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3781 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3841 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3901 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3961 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4021 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4081 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt cctgatgcgg
     4141 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     4201 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     4261 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     4321 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     4381 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     4441 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4501 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4561 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     4621 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     4681 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     4741 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     4801 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     4861 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     4921 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     4981 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5041 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5101 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     5161 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     5221 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5281 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5341 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     5401 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     5461 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5521 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5581 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5641 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5701 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     5761 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5821 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5881 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5941 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc
     6001 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6061 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6121 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6181 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6241 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     6301 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     6361 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     6421 agctgggctg caggctcaca tgt
//
