LOCUS       Exported                6469 bp ds-DNA     circular SYN 22-DEC-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    D434_pAAV-hSyn-SIO-GtACR1-TS-Fred-Kv2.1.dna
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6469)
  AUTHORS   Tivan27
  TITLE     Direct Submission
  JOURNAL   Exported Dec 22, 2017 from SnapGene 4.0.2
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6469
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     promoter        165..633
                     /label=hSyn1
     primer_bind     567..584
                     /label=OS09
                     /note="Sequences forward from Synapsin promoter"
     primer_bind     575..593
                     /label=OS08
                     /note="Sequences forward from CamKII promoter"
     primer_bind     585..604
                     /label=OS10
                     /note="Sequence from 3' of human synapsin promoter into 
                     insert"
     primer_bind     618..640
                     /label=OS25
                     /note="REV from end of hSyn promoter"
     protein_bind    647..680
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="loxP mutant. Cre-mediated recombination occurs in 
                     the 8-bp core sequence (GCATACAT)."
     CDS             complement(687..887)
                     /codon_start=1
                     /label=Kv2.1 targeting domain
                     /note="color: #993366"
                     /translation="QSQPILNTKEMAPQSKPPEELEMSSMPSPVAPLPARTEGVIDMRS
                     MSSIDSFISCATDFPEATRFK"
     primer_bind     complement(860..910)
                     /label=FRed-Kv2.1-fusion
     CDS             complement(888..1583)
                     /codon_start=1
                     /label=FusionRed
                     /translation="MVSELIKENMPMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFMYGSRTFIKHPPGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKVRGVNFPANGPVMQKKTLGWEASTETMYPADGGLEGACDMAL
                     KLVGGGHLICNLETTYRSKKPATNLKMPGVYNVDHRLERIKEADDETYVEQHEVAVARY
                     STGGAGDGGK"
     primer_bind     888..911
                     /label=TG_AgeI_FR_rev_(258)
                     /note="63,,"
     primer_bind     complement(932..950)
                     /label=seq_Fp635-C_For_(375)
                     /note="0.025,DST,"
     primer_bind     932..950
                     /label=seq_Fp635-C_For_(376)
                     /note="0.025,DST,"
     primer_bind     complement(1065..1082)
                     /label=lox_o_mKate(168)_for_(154)
                     /note="51,,"
     primer_bind     complement(1413..1425)
                     /label=ChR2_o_P2A_r_(112)
                     /note="57,,"
     primer_bind     complement(1413..1425)
                     /label=ChR2_o_PR_r_(113)
                     /note="54,,"
     primer_bind     complement(1430..1439)
                     /label=AscI_mRFP_rev_(251)
                     /note="47,,"
     primer_bind     complement(1430..1439)
                     /label=EcoRI_RFP_rev_(198)
                     /note="45,DST,"
     primer_bind     1496..1518
                     /label=seq_TurboBFP_rev_(346)_(246)
                     /note="0.025,DST,"
     primer_bind     complement(1557..1629)
                     /label=ts_FR_for_(424)
                     /note="0.05,DST,73"
     primer_bind     complement(1557..1583)
                     /label=AscI_FR_for_(257)
                     /note="48,,"
     primer_bind     complement(1560..1583)
                     /label=Primer 1
     primer_bind     complement(1560..1583)
                     /label=BamHI_FR_for_(351)
                     /note="0.025,DST,"
     primer_bind     complement(1560..1583)
                     /label=Fred_for_(388)
                     /note="0.025,DST,35"
     primer_bind     complement(1560..1583)
                     /label=Nhe_FRed_for_(353)
                     /note="0.025,DST,"
     primer_bind     complement(1563..1584)
                     /label=VVD_o_mKate2_for_(31)
                     /note="63,,"
     primer_bind     complement(1563..1583)
                     /label=PDFG_mKate_for_(18)
                     /note="71,,"
     primer_bind     complement(1572..1583)
                     /label=FRed_for_(381)
                     /note="0.025,DST,21"
     primer_bind     1574..1593
                     /label=FP635_for_(459)
                     /note="0.025,DST,"
     primer_bind     1584..1610
                     /label=P2A_C1V1_rev_(225)
                     /note="93,,"
     primer_bind     1584..1610
                     /label=P2A_C1V1_rev_(225) (2)
                     /note="93,,"
     primer_bind     1584..1599
                     /label=ts_rev_(372)
                     /note="0.025,DST,"
     CDS             complement(1587..1646)
                     /codon_start=1
                     /product="membrane trafficking sequence Kir2.1"
                     /label=ts
                     /translation="KSRITSEGEYIPLDQIDINV"
     primer_bind     1626..1670
                     /label=GtACR1-ts_fusion
     primer_bind     complement(1626..1644)
                     /label=seq_ts_for_(391)
                     /note="0.025,DST,19"
     primer_bind     complement(1626..1635)
                     /label=BeGC1_H564R_V568F_for_(475)
                     /note="0.025,DST,"
     primer_bind     1652..1665
                     /label=AgeI_GtACR1_rev_(480)
     CDS             complement(1653..2537)
                     /codon_start=1
                     /label=GtACR1
                     /translation="MSSITCDPAIYGEWSRENQFCVEKSLITLDGIKYVQLVMAVVSAC
                     QVFFMVTRAPKVPWEAIYLPTTEMITYSLAFTGNGYIRVANGKYLPWARMASWLCTCPI
                     MLGLVSNMALVKYKSIPLNPMMIAASSICTVFGITASVVLDPLHVWLYCFISSIFFIFE
                     MVVAFAIFAITIHDFQTIGSPMSLKVVERLKLMRIVFYVSWMAYPILWSFSSTGACIMS
                     ENTSSVLYLLGDALCKNTYGILLWATTWGLLNGKWDRDYVKGRNVDGTLMPEYEQDLEK
                     GNTERYEDARAGET"
     primer_bind     complement(2521..2549)
                     /label=BamHI_GtACR1_for_(479)
     primer_bind     complement(2523..2544)
                     /label=NheI_GtACR_for_(539)
     protein_bind    complement(2550..2583)
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="loxP mutant. Cre-mediated recombination occurs in 
                     the 8-bp core sequence (GCATACAT)."
     misc_feature    2596..3184
                     /label=WPRE
                     /note="woodchuck posttranscriptional regulatory element 
                     color: #ffffff"
     primer_bind     complement(2650..2671)
                     /label=OS07
                     /note="Sequences back from WPRE on Lenti vectors"
     CDS             complement(3067..3078)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3216..3692
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal color: 
                     #a6acb3"
     repeat_region   3732..3872
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      3947..4402
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis color: #ffff00"
     promoter        4684..4788
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             4789..5649
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics This feature has 2 segments:    1: 5567
                     ..   12 / #ccffcc / signal sequence    2:   13 ..  804 / 
                     #ccffcc Cleavage site after base 12"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5820..6408
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication color: #ffff00"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtgtgtc tagactgcag agggccctgc
      181 gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg
      241 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta
      301 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca
      361 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa
      421 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc
      481 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac
      541 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc
      601 agcggaggag tcgtgtcgtg cctgagagcg cagtcgagaa ggtacctacc gttcgtatag
      661 catacattat acgaagttat gaattcttac ttaaaccgcg tagcctctgg aaaatcggtg
      721 gcgcatgaga taaaagagtc aatagaactc atggacctca tgtcgatcac gccctctgtt
      781 cttgcaggaa gaggtgccac aggggagggc attgaggaca tttccagctc ctcagggggt
      841 ttactctggg gggccatctc tttggtgttc agaataggct gggactgttt acctccatca
      901 ccagcgccac cagtagagta tctggccaca gccacctcgt gctgctcgac gtatgtctca
      961 tcgtcggcct ccttgattct ttccagtctg tggtccacgt tgtagacgcc gggcatcttg
     1021 aggttcgtag cgggtttctt ggatctgtat gttgtctcaa ggttgcagat caggtgtcct
     1081 cctcccacga gcttcagagc catgtcacat gcgccttcca gtcctccgtc agcagggtac
     1141 atcgtctcgg tggaggcctc ccagccgagt gttttcttct gcatcacagg gccgttggct
     1201 gggaagttca cccctctaac cttgacgttg tagatgaggc agccgtcctg gaggctggtg
     1261 tcctgggtag cggtcagcac gcccccgtct tcgtatgtgg tgactctctc ccatgtgaag
     1321 ccctcaggga aggactgctt aaagaagtcg gggatgcccg gagggtgctt gatgaaggtt
     1381 ctgctgccgt acatgaagct ggtagccagg atgtcgaagg cgaaggggag agggccgccc
     1441 tcgacgacct tgattctcat ggtctgggtg ccctcgtagg gcttgccttc gccctcggat
     1501 gtgcacttga agtggtggtt gttcacggtg ccctccatgt acagcttcat gggcatgttc
     1561 tccttaatca gctcgctcac catcaccacg ttgatgtcga tctggtccag ggggatgtac
     1621 tcgccctcgc tggtgatcct gctcttggcg gcggtttccc cagcccgagc atcctcatat
     1681 cgttcagtat tgcctttctc cagatcctgt tcatattcgg gcatcagtgt cccatcgaca
     1741 ttgcgtcctt tcacgtaatc tcggtcccac ttgccgttca gcagtcccca ggtagtagcc
     1801 cacagcagga ttccataggt atttttgcac agtgcgtcgc ccagcaggta cagcacagag
     1861 gaagtgttct cagacataat gcaagcccct gtgcttgaaa aactccacag gataggatag
     1921 gccatccagc tcacgtagaa cacgattctc atcagtttca gcctttcgac caccttcagt
     1981 gacatgggac ttccgatggt ctgaaaatca tgaatagtga tggcaaaaat ggcgaatgcg
     2041 accaccatct caaagataaa gaaaatacta gagatgaaac agtacagcca cacgtgcagg
     2101 gggtccagga ccacggaggc agtgattccg aacactgtgc agatgctgct ggcggcaatc
     2161 atcatgggat tcagtggaat ggatttgtac ttcaccaggg ccatgttgct caccaggccc
     2221 agcatgattg gacatgtgca cagccaactt gccattctag cccaaggcag atacttccca
     2281 tttgccactc gaatgtatcc gttgcctgtg aaagccagtg aataggtgat catttctgtg
     2341 gttggcaggt agatggcctc ccaaggcact ttgggtgccc tagtgaccat aaagaacacc
     2401 tggcaggcgc tcaccacggc catgaccagc tgcacgtact tgatgccgtc cagggtaatc
     2461 aggctctttt cgacgcaaaa ctggttctct ctgctccact ctccatagat ggctgggtca
     2521 caagtaatgc tgctcatggt ggcggatcct accgttcgta taatgtatgc tatacgaagt
     2581 tataagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     2641 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     2701 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     2761 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     2821 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     2881 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     2941 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     3001 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     3061 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     3121 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     3181 ccgcatcgat accgagcgct gctcgagaga tctacgggtg gcatccctgt gacccctccc
     3241 cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa
     3301 aattaagttg catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg
     3361 gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt
     3421 gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg
     3481 ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca
     3541 ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg
     3601 tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac
     3661 aggcgtgaac cactgctccc ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac
     3721 cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     3781 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     3841 agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac
     3901 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
     3961 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4021 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4081 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4141 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     4201 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4261 actggaacaa cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg
     4321 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     4381 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
     4441 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     4501 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     4561 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     4621 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     4681 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4741 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     4801 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4861 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     4921 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     4981 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     5041 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     5101 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     5161 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     5221 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5281 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     5341 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     5401 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     5461 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5521 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     5581 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     5641 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     5701 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5761 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5821 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5881 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5941 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     6001 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6061 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6121 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6181 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6241 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6301 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6361 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6421 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
