LOCUS       Exported                5641 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Monomeric near-infrared fluorescent protein miRFP in 
            mammalian-expressing AAV production plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-Syn-miRFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5641)
  AUTHORS   Piatkevich KD, Jung EE, Straub C, Linghu C, Park D, Suk HJ, Hochbaum
            DR, Goodwin D, Pnevmatikakis E, Pak N, Kawashima T, Yang CT, Rhoades
            JL, Shemesh O, Asano S, Yoon YG, Freifeld L, Saulnier JL, Riegler C,
            Engert F, Hughes T, Drobizhev M, Szabo B, Ahrens MB, Flavell SW, 
            Sabatini BL, Boyden ES
  TITLE     A robotic multidimensional directed evolution approach applied to 
            fluorescent voltage reporters.
  JOURNAL   Nat Chem Biol. 2018 Feb 26. pii: 10.1038/s41589-018-0004-9. doi: 
            10.1038/s41589-018-0004-9.
  PUBMED    29483642
REFERENCE   2  (bases 1 to 5641)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5641
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        24..471
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      485..494
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    1457..2045
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1510..1530)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1928..1939)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2065..2541
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2264..2283)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2587..2727
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2587..2716
                     /label=AAV2 ITR
     rep_origin      2802..3257
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2889..2908)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3099..3120
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3274..3293)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3393..3415
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3453..3471)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3539..3643
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3644..4504
                     /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(3862..3881)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4675..5263
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5164..5183
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5325..5454
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gactgcagag ggccctgcgt atgagtgcaa gtgggtttta ggaccaggat gaggcggggt
       61 gggggtgcct acctgacgac cgaccccgac ccactggaca agcacccaac ccccattccc
      121 caaattgcgc atcccctatc agagaggggg aggggaaaca ggatgcggcg aggcgcgtgc
      181 gcactgccag cttcagcacc gcggacagtg ccttcgcccc cgcctggcgg cgcgcgccac
      241 cgccgcctca gcactgaagg cgcgctgacg tcactcgccg gtcccccgca aactcccctt
      301 cccggccacc ttggtcgcgt ccgcgccgcc gccggcccag ccggaccgca ccacgcgagg
      361 cgcgagatag gggggcacgg gcgcgaccat ctgcgctgcg gcgccggcga ctcagcgctg
      421 cctcagtctg cggtgggcag cggaggagtc gtgtcgtgcc tgagagcgca gtcgagtagg
      481 atccgccacc atggtcgcag gacacgcaag tggaagcccc gactttggaa ccgccgatcc
      541 gagcgactgt gagagagaag aaatccatct ggccggatcc atccagccac acggaacact
      601 gctggtggtc tccgagcctg accatagaat cattcaggca tctgccaacg ccgctgaatt
      661 cctgaatctg ggaagtgtgc tgggagtccc cctggctgag atcgacgggg atctgctgat
      721 caagattctg ccccacctgg accctaccgc agaaggaatg ccagtggccg tcaggtgccg
      781 cattggaaac ccctccacag agtacgatgg actgatgcat cgaccacctg aaggaggact
      841 gatcattgag ctggaacggg ctgggccacc cattgatctg agcggaacac tggctcccgc
      901 actggagcga attcgaactg caggatccct gagggctctg tgcgacgata ctgcactgct
      961 gttccagcag tgtaccggct acgaccgagt gatggtctat cggttcgatg aacaggggca
     1021 cggagaggtg tattcagaaa tacatgtgac tggactggag agctactttg gcaacaggta
     1081 tcctagctcc ctcgtgccac agatggcccg gagactgtac gaaagacaga gggtgcgcgt
     1141 cctggtggat gtctcttatc agcctgtgcc actggagcca agactgagtc ccctgactgg
     1201 cagggacctg gatatgtcag ggtgttttct gcgctctatg agtcctactc acctgcagtt
     1261 cctgaagaat atgggcgtgc gggcaacact ggtggtcagc ctggtggtcg gaggcaaact
     1321 gtgggggctg gtgatctgcc accattatct gccacgcttc atccattttg agctgcgagc
     1381 aatttgtgag ctgctggccg aagctatcgc aacccgcatt acagccctgt aagaattccg
     1441 tacaaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     1501 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     1561 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     1621 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     1681 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     1741 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     1801 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     1861 ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt ccttctgcta
     1921 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     1981 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     2041 cccgcatcga taccggctcg agacgggtgg catccctgtg acccctcccc agtgcctctc
     2101 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc
     2161 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg
     2221 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc
     2281 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt
     2341 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa
     2401 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc
     2461 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc
     2521 actgctccct tccctgtcct tctcgagctg attttgtagg taaccacgtg cggaccgagc
     2581 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     2641 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     2701 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     2761 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     2821 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     2881 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     2941 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3001 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3061 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3121 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     3181 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     3241 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     3301 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     3361 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     3421 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     3481 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     3541 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3601 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     3661 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3721 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     3781 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     3841 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     3901 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     3961 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4021 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4081 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4141 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4201 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4261 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4321 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4381 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4441 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4501 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     4561 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4621 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4681 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4741 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4801 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     4861 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4921 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4981 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5041 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5101 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5161 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5221 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5281 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtcctgca ggcagctgcg
     5341 cgctcgctcg ctcactgagg ccgcccgggc gtcgggcgac ctttggtcgc ccggcctcag
     5401 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc
     5461 gcacgcgttg caaagatgga taaagtttta aacagagagg aatctttgca gctaatggac
     5521 cttctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg cagagcgcac
     5581 atcgcccaca gtccccgaga agttgggggg aggggtcggc agcaaatggt taattaatct
     5641 a
//
