LOCUS       Exported                6017 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresssion of Ilmol preferentially in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    AAV_hSyn_Ilmol
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6017)
  AUTHORS   Yi B, Kang BE, Lee S, Braubach S, Baker BJ
  TITLE     A dimeric fluorescent protein yields a bright, red-shifted GEVI 
            capable of population signals in brain slice.
  JOURNAL   Sci Rep. 2018 Oct 12;8(1):15199. doi: 10.1038/s41598-018-33297-y.
  PUBMED    30315245
REFERENCE   2  (bases 1 to 6017)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6017
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   6..135
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        180..627
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             1420..2121
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLRGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(1553..1571)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1821..1840
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2015..2038
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    2149..2737
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2202..2222)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2620..2631)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2769..3245
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2968..2987)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3285..3425
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3285..3414
                     /label=AAV2 ITR
     rep_origin      3500..3955
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3587..3606)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3797..3818
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3972..3991)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4091..4113
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4151..4169)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4237..4341
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4342..5202
                     /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(4560..4579)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5373..5961
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5862..5881
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga
       61 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
      121 tcactagggg ttcctgcggc cgcacgcgtg tgtctagact gcagagggcc ctgcgtatga
      181 gtgcaagtgg gttttaggac caggatgagg cggggtgggg gtgcctacct gacgaccgac
      241 cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag
      301 agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg
      361 acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg
      421 ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc
      481 gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc
      541 gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga
      601 ggagtcgtgt cgtgcctgag agcgcagtcg agaaggtacc ggatccgcca ccggagaccc
      661 aagctggcta gcaccatgga gggattcgac ggttcagatt ttagtcctcc agctgattta
      721 gttggcgttg acggtgcagt catgcggaac gtcgttgacg tcacgataaa tggtgacgtc
      781 actgctccgc cgaaagctgc gccaagaaaa agtgaatcgg taaagaaagt tcattggaat
      841 gatgtagacc aaggaccgag tgaaaaacca gagacaagac aggaggaacg aatagatata
      901 cccgagattt caggtctatg gtggggcgag aatgaacatg gagtggacga tgggagaatg
      961 gagataccta ctactggtgt aggtcgcgtc cagtttcgca ttcgagcagt gattgatcat
     1021 ctagggatgc gagtctttgg agtcttccta attttcttgg acatcatcct catgatcatt
     1081 gatctcagtc ttatcgataa aagtgaatct tcacaatcct tttatgacgg gatggacttg
     1141 gctctttctt gttatttcat gctggattta ggattaagga tatttgccta cgggcccaag
     1201 cttttcttca ccaacccttg ggaggtggct gatggtttga ttatcgttgt cacattcgtc
     1261 gtcacgatat tttacactgt gttagatgaa tacggtaccg aaacaggagc cgatggtttg
     1321 gggcagttgg ttgtgttggc ccgtttgctg cgtgtggtta tcttagcaag aatattttat
     1381 tccagacaac aaatgaaggc ttcaagcaga ggggatccca tggtgagcaa gggcgaggag
     1441 gtcatcaaag agttcatgcg cttcaaggtg cgcatggagg gctccatgaa cggccacgag
     1501 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     1561 aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc ccagttcatg
     1621 tacggctcca aggcgtacgt gaagcacccc gccgacatcc ccgattacaa gaagctgtcc
     1681 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg tctggtgacc
     1741 gtgacccagg actcctccct gcaggacggc acgctgatct acaaggtgaa gatgcgcggc
     1801 accaacttcc cccccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     1861 accgagcgcc tgtacccccg cgacggcgtg ctgagaggcg agatccacca ggccctgaag
     1921 ctgaaggacg gcggccacta cctggtggag ttcaagacca tctacatggc caagaagccc
     1981 gtgcaactgc ccggatacta ctacgtggac accaagctgg acatcacctc ccacaacgag
     2041 gactacacca tcgtggaaca gtacgagcgc tccgagggcc gccaccacct gttcctgtac
     2101 ggcatggacg agctgtacaa gtgagaattc gatatcaagc ttatcgataa tcaacctctg
     2161 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     2221 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     2281 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     2341 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     2401 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     2461 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     2521 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc
     2581 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     2641 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     2701 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag
     2761 agatctacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg
     2821 ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact
     2881 aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg
     2941 ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac
     3001 aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc
     3061 ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt
     3121 agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct
     3181 acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg
     3241 tccttctgat tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat
     3301 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     3361 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     3421 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     3481 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3541 gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc
     3601 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3661 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     3721 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3781 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg
     3841 ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag
     3901 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     3961 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     4021 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4081 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     4141 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     4201 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4261 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4321 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4381 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4441 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4501 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     4561 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     4621 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     4681 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     4741 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     4801 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     4861 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     4921 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     4981 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5041 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5101 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5161 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     5221 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5281 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5341 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5401 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5461 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5521 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5581 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5641 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5701 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5761 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5821 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5881 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5941 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6001 tgctggcctt ttgctca
//
