LOCUS       Exported                7318 bp ds-DNA     circular SYN 24-AUG-2020
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    p131_pAAV-EF1a-NES(PKIt)-tdTomato-biLINuS10-WPRE
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7318)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Monday, August 24, 2020 from SnapGene 3.3.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7318
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
                     /note="color: #ffe4c4"
     promoter        242..1419
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
                     /note="color: #ffffff; direction: RIGHT"
     intron          473..1410
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
                     /note="color: #------"
     CDS             1438..3420
                     /codon_start=1
                     /label=NES(PKIt)-tdTomato-biNLS10
                     /note="color: #993366"
                     /translation="MLALKLAGLDIASMVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEG
                     EGEGRPYEGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEG
                     FKWERVMNFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTER
                     LYPRDGVLKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDY
                     TIVEQYERSEGRHHLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGH
                     EFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKK
                     LSFPEGFKWERVMNFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGW
                     EASTERLYPRDGVLKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDIT
                     SHNEDYTIVEQYERSEGRHHLFLYGMDELYKGGSGGSGGSLATTLERIEKNFVITDPRL
                     PDNPIIFASDSFLQLTEYSREEILGRNCRFLQGPETDRATVRKIRDAIDNQTEVTVQLI
                     NYTKSGKKFWNLFHLQPMRDQKGDVQYFIGVQLDGTEHVRDAAEREGVMLIKKTAENID
                     EAAKRLPDANLAAAAAAAAKKKKLD"
     CDS             1441..1470
                     /codon_start=1
                     /label=NES(PKIt)
                     /note="color: #993366"
                     /translation="LALKLAGLDI"
     CDS             1477..2904
                     /codon_start=1
                     /label=tdTomato
                     /note="color: #ff0000"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     CDS             2932..3417
                     /codon_start=1
                     /label=biLINuS10
                     /note="color: #993366"
                     /translation="LATTLERIEKNFVITDPRLPDNPIIFASDSFLQLTEYSREEILGR
                     NCRFLQGPETDRATVRKIRDAIDNQTEVTVQLINYTKSGKKFWNLFHLQPMRDQKGDVQ
                     YFIGVQLDGTEHVRDAAEREGVMLIKKTAENIDEAAKRLPDANLAAAAAAAAKKKKLD"
     misc_feature    3445..4033
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
                     /note="color: #ffffff"
     terminator      4063..4541
                     /label=hGH_PA_ terminator
                     /label=hGH_PA_terminator
                     /note="color: #ffffff"
     repeat_region   4581..4721
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
                     /note="color: #ffe4c4"
     rep_origin      4796..5251
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     misc_feature    5387..5409
                     /label=pGEX_3_primer
                     /note="color: #a6acb3"
     promoter        5533..5637
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     promoter        5568..5596
                     /label=AmpR_ promoter
                     /label=AmpR_promoter
                     /note="color: #ffffff"
     gene            5638..6498
                     /gene="Ampicillin"
                     /label=Ampicillin
                     /note="color: #a6acb3; direction: RIGHT"
     CDS             5638..6498
                     /codon_start=1
                     /label=ORF frame 1
                     /note="color: #993366"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6669..7257
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc cgccaccatg
     1441 ttagccttga aattagcagg tcttgatatc gctagcatgg tgagcaaggg cgaggaggtc
     1501 atcaaagagt tcatgcgctt caaggtgcgc atggagggct ccatgaacgg ccacgagttc
     1561 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
     1621 gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccca gttcatgtac
     1681 ggctccaagg cgtacgtgaa gcaccccgcc gacatccccg attacaagaa gctgtccttc
     1741 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggtct ggtgaccgtg
     1801 acccaggact cctccctgca ggacggcacg ctgatctaca aggtgaagat gcgcggcacc
     1861 aacttccccc ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctccacc
     1921 gagcgcctgt acccccgcga cggcgtgctg aagggcgaga tccaccaggc cctgaagctg
     1981 aaggacggcg gccactacct ggtggagttc aagaccatct acatggccaa gaagcccgtg
     2041 caactgcccg gctactacta cgtggacacc aagctggaca tcacctccca caacgaggac
     2101 tacaccatcg tggaacagta cgagcgctcc gagggccgcc accacctgtt cctggggcat
     2161 ggcaccggca gcaccggcag cggcagctcc ggcaccgcct cctccgagga caacaacatg
     2221 gccgtcatca aagagttcat gcgcttcaag gtgcgcatgg agggctccat gaacggccac
     2281 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
     2341 ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc cccccagttc
     2401 atgtacggct ccaaggcgta cgtgaagcac cccgccgaca tccccgatta caagaagctg
     2461 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggtctggtg
     2521 accgtgaccc aggactcctc cctgcaggac ggcacgctga tctacaaggt gaagatgcgc
     2581 ggcaccaact tcccccccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
     2641 tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca ccaggccctg
     2701 aagctgaagg acggcggcca ctacctggtg gagttcaaga ccatctacat ggccaagaag
     2761 cccgtgcaac tgcccggcta ctactacgtg gacaccaagc tggacatcac ctcccacaac
     2821 gaggactaca ccatcgtgga acagtacgag cgctccgagg gccgccacca cctgttcctg
     2881 tacggcatgg acgagctgta caagggtgga tctggaggtt caggtggaag tttggctact
     2941 acacttgaac gtattgagaa gaactttgtc attactgacc caagattgcc agataatccc
     3001 attatattcg cgtccgatag tttcttgcag ttgacagaat atagccgtga agaaattttg
     3061 ggaagaaact gcaggtttct acaaggtcct gaaactgatc gcgcgacagt gagaaaaatt
     3121 agagatgcca tagataacca aacagaggtc actgttcagc tgattaatta tacaaagagt
     3181 ggtaaaaagt tctggaacct ctttcacttg cagcctatgc gagatcagaa gggagatgtc
     3241 cagtacttta ttggggttca gttggatgga actgagcatg tccgagatgc tgccgagaga
     3301 gagggagtca tgctgattaa gaaaactgca gaaaatattg acgaggccgc aaagagactg
     3361 cccgacgcca acctggccgc agcagccgca gccgcagcaa agaagaaaaa gctggactag
     3421 gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     3481 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     3541 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     3601 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     3661 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     3721 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     3781 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     3841 tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc
     3901 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     3961 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     4021 gccgcctccc cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg catccctgtg
     4081 acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg
     4141 tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg
     4201 ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg
     4261 gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg
     4321 cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat
     4381 gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg
     4441 ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc
     4501 tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca
     4561 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     4621 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     4681 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     4741 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     4801 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4861 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4921 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4981 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     5041 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     5101 ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga tttataaggg
     5161 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     5221 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     5281 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     5341 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     5401 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     5461 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     5521 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5581 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5641 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5701 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5761 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5821 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5881 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5941 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     6001 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6061 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6121 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     6181 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6241 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6301 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6361 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6421 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6481 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6541 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6601 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6661 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6721 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6781 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     6841 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6901 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6961 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7021 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7081 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7141 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7201 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7261 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
