LOCUS       Exported                6398 bp ds-DNA     linear   SYN 26-MAY-2021
DEFINITION  Expresses Jaws-mRuby2 fusion in GABAergic inhibitory neurons in 
            rodent cortex.
ACCESSION   .
VERSION     .
KEYWORDS    AAV-hSyn-Jaws-mRuby2-8X2C
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6398)
  AUTHORS   Keaveney MK, Tseng HA, Ta TL, Gritton HJ, Man HY, Han X
  TITLE     A MicroRNA-Based Gene-Targeting Tool for Virally Labeling 
            Interneurons in the Rodent Cortex.
  JOURNAL   Cell Rep. 2018 Jul 10;24(2):294-303. doi: 
            10.1016/j.celrep.2018.06.049.
  PUBMED    29996091
REFERENCE   2  (bases 1 to 6398)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 26, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6398
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      71..526
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(158..177)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     368..389
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        552..656
                     /gene="bla"
                     /label=AmpR promoter
     CDS             657..1517
                     /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(875..894)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1688..2276
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2177..2196
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2430..2447
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2564..2585
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2600..2630
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2638..2654
                     /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     2643..2665
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2662..2678
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2662..2678
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   2692..2821
                     /label=AAV2 ITR
     promoter        2915..3362
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             4221..4928
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="VSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTMR
                     IKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGGV
                     VTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTHM
                     ALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAVA
                     KFAGLGGGMDELYK"
     misc_feature    5459..6047
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5512..5532)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5930..5941)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6050..6066)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6051..6067)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    6096..6217
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6107..6126)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6161..6180
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
       61 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
      121 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
      181 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
      241 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
      301 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
      361 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
      421 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
      481 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
      541 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
      601 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
      661 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
      721 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
      781 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
      841 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
      901 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
      961 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     1021 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     1081 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     1141 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     1201 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     1261 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     1321 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     1381 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     1441 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     1501 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     1561 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     1621 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     1681 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     1741 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     1801 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     1861 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     1921 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     1981 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     2041 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     2101 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     2161 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     2221 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     2281 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     2341 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     2401 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     2461 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     2521 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     2581 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     2641 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc actgcgcgct
     2701 cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg
     2761 gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc
     2821 ttgtagttaa tgattaaccc gccatgctac ttatctacgt agccatgctc taggaagatc
     2881 tgaattctct agactgcaga gggccctgcg tatgagtgca agtgggtttt aggaccagga
     2941 tgaggcgggg tgggggtgcc tacctgacga ccgaccccga cccactggac aagcacccaa
     3001 cccccattcc ccaaattgcg catcccctat cagagagggg gaggggaaac aggatgcggc
     3061 gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt gccttcgccc ccgcctggcg
     3121 gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac gtcactcgcc ggtcccccgc
     3181 aaactcccct tcccggccac cttggtcgcg tccgcgccgc cgccggccca gccggaccgc
     3241 accacgcgag gcgcgagata ggggggcacg ggcgcgacca tctgcgctgc ggcgccggcg
     3301 actcagcgct gcctcagtct gcggtgggca gcggaggagt cgtgtcgtgc ctgagagcgc
     3361 agtcgagagg atccgccgcc accatgaccg ccgtgagcac cacagccact accgtgctgc
     3421 aggccacaca gagcgacgtg ctgcaggaga tccagtccaa cttcctgctg aatagctcca
     3481 tctgggtgaa cattgctctg gccggagtgg tcatcctgct gtttgtggcc atggggaggg
     3541 atctggaatc ccctagagct aagctgatct gggtggccac aatgctggtg ccactggtgt
     3601 ctatttctag ttacgctgga ctggccagtg ggctgactgt gggcttcctg cagatgccac
     3661 ctggacacgc tctggccgga caggaggtgc tgagcccatg gggccggtat ctgacatgga
     3721 ctttctccac tcccatgatc ctgctggctc tgggactgct ggccgacacc gatattgcca
     3781 gcctgttcac cgccatcaca atggacattg gcatgtgcgt gacaggactg gccgctgccc
     3841 tgatcactag ctcccatctg ctgcgctggg tgttctacgg aatttcttgt gctttctttg
     3901 tggccgtgct gtatgtgctg ctggtgcagt ggccagctga tgctgaggct gctgggacca
     3961 gtgaaatctt tggcactctg aggattctga ccgtggtgct gtggctgggg taccctatcc
     4021 tgttcgctct gggctctgag ggagtggccc tgctgagtgt gggagtgacc agctggggat
     4081 actccggact ggacatcctg gctaaatacg tgttcgcctt tctgctgctg agatgggtgg
     4141 ctgccaatga aggcacagtg tctgggagtg gaatgggaat cgggtccgga ggagctgctc
     4201 cagccgacga tcgaccggta gtgtctaagg gcgaagagct gatcaaggaa aatatgcgta
     4261 tgaaggtggt catggaaggt tcggtcaacg gccaccaatt caaatgcaca ggtgaaggag
     4321 aaggcaatcc gtacatggga actcaaacca tgaggatcaa agtcatcgag ggaggacccc
     4381 tgccatttgc ctttgacatt cttgccacgt cgttcatgta tggcagccgt acttttatca
     4441 agtacccgaa aggcattcct gatttcttta aacagtcctt tcctgagggt tttacttggg
     4501 aaagagttac gagatacgaa gatggtggag tcgtcaccgt catgcaggac accagccttg
     4561 aggatggctg tctcgtttac cacgtccaag tcagaggggt aaactttccc tccaatggtc
     4621 ccgtgatgca gaagaagacc aagggttggg agcctaatac agagatgatg tatccagcag
     4681 atggtggtct gaggggatac actcatatgg cactgaaagt tgatggtggt ggccatctgt
     4741 cttgctcttt cgtaacaact tacaggtcaa aaaagaccgt cgggaacatc aagatgcccg
     4801 gtatccatgc cgttgatcac cgcctggaaa ggttagagga aagtgacaat gaaatgttcg
     4861 tagtacaacg cgaacacgca gttgccaagt tcgccgggct tggtggtggg atggacgagc
     4921 tgtacaagct gtacaagttc tgctacgaga atgaagtgta aactagtgcg gccttaaaga
     4981 gaccggttca ctgtgacagt aaaagagacc ggttcactgt gagaatgaaa gagaccggtt
     5041 cactgtgatc ggaaaagaga ccggttcact gtgagcggcc ttaaagagac cggttcactg
     5101 tgacagtaaa agagaccggt tcactgtgag aatgaaagag accggttcac tgtgatcgga
     5161 aaagagaccg gttcactgtg agcggccttg aaacccagca gacaatgtag ctcagtagaa
     5221 acccagcaga caatgtagct gaatggaaac ccagcagaca atgtagcttc ggagaaaccc
     5281 agcagacaat gtagctgcgg ccttgaaacc cagcagacaa tgtagctcag tagaaaccca
     5341 gcagacaatg tagctgaatg gaaacccagc agacaatgta gcttcggaga aacccagcag
     5401 acaatgtagc tgcggccgcg actctagagt cgacctgcag gcatgcaagc ttatcgataa
     5461 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     5521 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     5581 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     5641 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     5701 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     5761 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     5821 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     5881 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     5941 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     6001 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg
     6061 acctcgaccc gggcggccgc ttcgagcaga catgataaga tacattgatg agtttggaca
     6121 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     6181 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     6241 tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa acctctacaa
     6301 atgtggtaaa atcgataagg atcttcctag agcatggcta cgtagataag tagcatggcg
     6361 ggttaatcat taactacaag gaacccctag tgatggag
//
