LOCUS       Exported                6292 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses iAChSnFR under GFAP promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV.ZAC.GFAP.iAChSnFR
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6292)
  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  JOURNAL   Unpublished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 6292)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6292
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    125..146
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        161..191
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    199..215
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     204..226
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     223..239
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     223..239
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   277..406
                     /label=AAV2 ITR
     intron          1257..1389
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1372..1391
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1434..1452
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     1469..1531
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
     primer_bind     1981..2002
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2105..2126)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             3113..3142
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3146..3292
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     polyA_signal    3335..3456
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3346..3365)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3400..3419
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3618..3747
                     /label=AAV2 ITR
     primer_bind     complement(3766..3783)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3766..3782)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3775..3797)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3924..4379
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4011..4030)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4221..4242
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4405..4509
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4510..5370
                     /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(4728..4747)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5541..6129
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6030..6049
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6283..8
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
       61 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
      121 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
      181 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
      241 atgattacgc cagatttaat taaggcctta attaggctgc gcgctcgctc gctcactgag
      301 gccgcccggg caaagcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag
      361 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttccttgta gttaatgatt
      421 aacccgccat gctacttatc tacgtagcca tgctctagga agatctaaca tatcctggtg
      481 tggagtaggg gacgctgctc tgacagaggc tcgggggcct gagctggctc tgtgagctgg
      541 ggaggaggca gacagccagg ccttgtctgc aagcagacct ggcagcattg ggctggccgc
      601 cccccagggc ctcctcttca tgcccagtga atgactcacc ttggcacaga cacaatgttc
      661 ggggtgggca cagtgcctgc ttcccgccgc accccagccc ccctcaaatg ccttccgaga
      721 agcccattga gcagggggct tgcattgcac cccagcctga cagcctggca tcttgggata
      781 aaagcagcac agccccctag gggctgccct tgctgtgtgg cgccaccggc ggtggagaac
      841 aaggctctat tcagcctgtg cccaggaaag gggatcaggg gatgcccagg catggacagt
      901 gggtggcagg gggggagagg agggctgtct gcttcccaga agtccaagga cacaaatggg
      961 tgaggggaga gctctcccca tagctgggct gcggcccaac cccaccccct caggctatgc
     1021 cagggggtgt tgccaggggc acccgggcat cgccagtcta gcccactcct tcataaagcc
     1081 ctcgcatccc aggagcgagc agagccagag caggttggag aggagacgca tcacctccgc
     1141 tgctcgcaag ctttattgcg gtagtttatc acagttaaat tgctaacgca gtcagtgctt
     1201 ctgacacaac agtctcgaac ttaagctgca gaagttggtc gtgaggcact gggcaggtaa
     1261 gtatcaaggt tacaagacag gtttaaggag accaatagaa actgggcttg tcgagacaga
     1321 gaagactctt gcgtttctga taggcaccta ttggtcttac tgacatccac tttgcctttc
     1381 tctccacagg tgtccactcc cagttcaatt acagctctta aggctagagt acttaatacg
     1441 actcactata ggctagcctc gagaattcat ggagacagac acactcctgc tatgggtact
     1501 gctgctctgg gttccaggtt ccactggtga cagatctgcg aacgacaccg tagttgtggg
     1561 ctcgatcatc tttacagaag ggattatcgt cgcaaacatg gtggcagaga tgattgaggc
     1621 gcatacagac cttaaggtgg ttcgcaaact gaaccttggc ggggtcaacg ttaactttga
     1681 agccattaaa cgcggaggtg cgaataatgg tattgacatt tacgtggagt acactgggca
     1741 cggtcttgtg gatattctgg ggttcccgga gccgaacgtc tatatcaccg ccgacaagca
     1801 gaagaacggc atcaaggcga acttcaagat ccgccacaac gtggaggacg gcagcgtgca
     1861 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1921 caaccactac ctgagcaccc agtccgtgct gagcaaagac cccaacgaga agcgcgatca
     1981 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     2041 caagggcggt accggaggga gcatgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     2101 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgc gcggcgaggg
     2161 cgagggcgat gccaccaacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     2221 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     2281 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     2341 ccaggagcgc accatcagct tcaaggacga cggcacctac aagacccgcg ccgaggtgaa
     2401 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     2461 cggcaacatc ctggggcaca agctggagta caactttccg ccgcccgcga ctactgatcc
     2521 agaaggtgca tacgaaaccg tgaagaagga gtacaaacgt aaatggaata ttgtatggct
     2581 caaaccactg ggattcaaca atacgtatac gcttaccgtt aaagacgaac tggcgaaaca
     2641 gtataacctt aaaaccttca gtgacttagc gaaaatctcg gataagctga ttctgggtgc
     2701 aacgatgttc tttttagaag ggcccgatgg ttacccaggc ctgcaaaaac tgtacaattt
     2761 caaattcaag cacaccaaaa gcatggacat gggtattcgc tataccgcca ttgataataa
     2821 cgaagttcag gtaattgatg cctgggccac tgatggcttg ctggtgagcc acaaattaaa
     2881 aattctggag gatgataaag cgttcttccc gccgtattat gctgccccca tcatccgtca
     2941 ggatgtctta gataagcatc ctgaactgaa ggacgtgctg aacaaactcg cgaatcaaat
     3001 ttcactggaa gaaatgcaga aactgaatta caaggtggac ggtgagggtc aggacccagc
     3061 gaaagtagct aaggagtttt tgaaagagaa aggtttaatt ctgcaggtcg acgaacaaaa
     3121 actcatctca gaagaggatc tgaatgctgt gggccaggac acgcaggagg tcatcgtggt
     3181 gccacactcc ttgcccttta aggtggtggt gatctcagcc atcctggccc tggtggtgct
     3241 caccatcatc tcccttatca tcctcatcat gctttggcag aagaagccac gttagtctag
     3301 agtcgacccg ggcggccgct tcgagcagac atgataagat acattgatga gtttggacaa
     3361 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     3421 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     3481 atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     3541 tgtggtaaaa tcgataagga tcttcctaga gcatggctac gtagataagt agcatggcgg
     3601 gttaatcatt aactacaagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     3661 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     3721 ggcctcagtg agcgagcgag cgcgcagcct taattaacct aattcactgg ccgtcgtttt
     3781 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     3841 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     3901 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3961 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4021 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4081 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4141 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4201 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4261 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4321 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     4381 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4441 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4501 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4561 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4621 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4681 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4741 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4801 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4861 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4921 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4981 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5041 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5101 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5161 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5221 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5281 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5341 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5401 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5461 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5521 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5581 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5641 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5701 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5761 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5821 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5881 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5941 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6001 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6061 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6121 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6181 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     6241 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc ag
//
