LOCUS       Exported                6782 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Fluorescent reporter for serotonin (dendrite localization tag).
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-CAG-iSeroSnFR-Nlgn
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6782)
  AUTHORS   Unger EK, Keller JP, Altermatt M, Liang R, Matsui A, Dong C, Hon OJ,
            Yao Z, Sun J, Banala S, Flanigan ME, Jaffe DA, Hartanto S, Carlen J,
            Mizuno GO, Borden PM, Shivange AV, Cameron LP, Sinning S, Underhill 
            SM, Olson DE, Amara SG, Temple Lang D, Rudnick G, Marvin JS, Lavis 
            LD, Lester HA, Alvarez VA, Fisher AJ, Prescher JA, Kash TL, 
            Yarov-Yarovoy V, Gradinaru V, Looger LL, Tian L
  TITLE     Directed Evolution of a Selective and Sensitive Serotonin Sensor via
            Machine Learning.
  JOURNAL   Cell. 2020 Dec 12. pii: S0092-8674(20)31612-3. doi: 
            10.1016/j.cell.2020.11.040.
  PUBMED    33333022
REFERENCE   2  (bases 1 to 6782)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6782
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..278
                     /label=chicken beta-actin promoter
     primer_bind     557..579
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     641..660
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      696..705
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     1295..1316
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(2688..2699)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    3105..3693
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3158..3178)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3576..3587)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3736..3857
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3747..3766)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3801..3820
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4192..4209)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4363..4951)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4443..4462)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5122..5982)
                     /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     5745..5764
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5983..6087)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6113..6568)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6250..6271)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6462..6481
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa
       61 ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg
      121 ggcgcgcgcc gggggggggg gggggggggg gggggsgggg sggggsgagg cggagaggtg
      181 cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc
      241 ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc
      301 gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt
      361 tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg
      421 tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg
      481 gccctttgtg cggggggagc ggctcggggc tgtccgcggg gggacggctg ccttcggggg
      541 ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag agcctctgct
      601 aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg
      661 ctgtctcatc attttggcaa agaattggat ctgccgccac catggcactt cctagatgta
      721 tgtggcctaa ttatgtatgg cgagctatga tggcctgcgt ggtgcaccga gggagtggcg
      781 ccccacttac actctgcctg ttgggttgcc tcctccagac tttccacgtt ctttcacaga
      841 agctggaatt cgcgaacgac accgtagttg tgggctcgat taaccataca gaacagatta
      901 tcgtcgcaaa catgttggca gagatgattg aggcgcatac agaccttaag gtggttcgca
      961 aactgaacct tggcggggtt aacgttaact ttgaagccat taaacgcgga ggtgcgaata
     1021 atggtattga catttacctg gagtacgttg ggtatggtct tgtggatatt ctggggtacc
     1081 cggagccgaa cgtctatatc atcgccgaca agcagaagaa cggcatcaag gcgaacttca
     1141 agatccgcta caacgtggag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     1201 cccccatcgg cgacggccct gtgctgctgc ctgacaacca ctacctgagc acccagtccg
     1261 tgctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     1321 ccgccgggat cactctcggc atggacgagc tgtacaaggg cggtaccgga gggagcatga
     1381 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcggcg
     1441 taaacggcca caagttcagc gtgcgcggcg agggcgaggg cgatgccacc aacggcaagc
     1501 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     1561 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     1621 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc agcttcaagg
     1681 acgacggcac ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     1741 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     1801 agtacaactt tccgccgccc ggtgctactg atccagaagg tgcatacgaa accgtgaaga
     1861 aggagtacaa acgtaaatgg aatattgtat ggctcaaacc actgggattc aacgcttcgt
     1921 atgtgcttgc ggttaaagac gaactggcga aacagtataa ccttaaaacc ttcagtgact
     1981 tagcgaaaat ctcggataag ctgattctgg gtgcaaacat gatgttttta gaaaatcccg
     2041 atggttaccc aggcctgcaa aaactgtaca atttcaaatt caagcacacc aaaagcatgg
     2101 acgctggtat tccgtatacc gccattgata ataacgaagt tcaggtaatt gatgccactg
     2161 ccactgatgg cttgctggtg agccacaaat taaaaattct ggaggatgat aaagcgttct
     2221 tcccgccgta ttatgctgcc cccatcatcc gtcaggatgt cttagataag catcctgaac
     2281 tgaaggacgt gctgaacaaa ctcgcgaatc aaatttcact ggaagaaatg cagaaactga
     2341 attacaagcg tgacggtgag ggtcaggacc cagcgaaagt agctaaggag tttttgaaag
     2401 agaaaggttt aattggatcc acgctcgagc tggtgcccca cttgcataac ctcaatgaca
     2461 tcagccaata cacaagtaca accaccaagg tcccaagcac tgacatcacc cttcgaccaa
     2521 ctcggaaaaa ctcaacccct gtgaccagcg catttccaac cgctaagcaa gacgacccca
     2581 aacaacagcc tagtccattt agcgtagacc aaagggacta cagcacagag ctgtccgtga
     2641 ccatagctgt aggagcttcc ctgcttttcc tgaacattct ggctttcgcg gccctctatt
     2701 acaagaagga caaaaggcgc cacgacgtac ataggagatg tagtccgcaa cgaactacaa
     2761 ccaatgatct tacgcacgca cctgaggagg agatcatgag tcttcagatg aaacacacag
     2821 acctggacca cgagtgtgaa tccatccatc cccatgaggt ggtgctgagg acagcatgcc
     2881 ctccggacta taccctggca atgcgccgaa gcccagacga cattcccctg atgacaccca
     2941 ataccataac catgattccc aacactattc caggaatcca gcctctccac actttcaaca
     3001 cattcactgg tggacagaat aatactctcc ctcatccgca tccacacccc cactcccaca
     3061 gcacaacacg ggtgggtggc agcggtggct aaaagcttat cgataatcaa cctctggatt
     3121 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     3181 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     3241 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     3301 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     3361 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     3421 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     3481 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
     3541 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     3601 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     3661 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgaccc gggcggccgc
     3721 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     3781 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     3841 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     3901 agatgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg
     3961 atcttcctag agcatggcta cgtagataag tagcatggcg ggttaatcat taactacaag
     4021 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     4081 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc attaatgaat
     4141 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4201 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4261 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4321 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4381 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4441 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4501 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4561 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4621 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4681 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4741 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4801 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4861 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4921 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4981 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5041 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5101 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5161 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5221 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5281 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5341 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5401 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5461 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5521 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5581 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5641 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5701 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5761 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5821 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5881 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5941 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6001 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6061 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta
     6121 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     6181 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     6241 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     6301 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     6361 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     6421 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     6481 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     6541 gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg
     6601 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tgcgcgctcg ctcgctcact
     6661 gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc
     6721 gasckwrygs gmmkwkmskr mktggcagta catctacgta ttagtcatcg ctattaccat
     6781 gg
//
