LOCUS       Exported                7690 bp ds-DNA     circular SYN 04-MAY-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7690)
  AUTHORS   Adesnik Lab
  TITLE     Direct Submission
  JOURNAL   Exported May 4, 2021 from SnapGene 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7690
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    378..509
                     /label=5'ITR
                     /note="color: #00ffff"
     promoter        823..1100
                     /label=chicken beta-actin promoter
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    1511..1515
                     /label=BamHI
                     /note="color: #f5ff7b"
     misc_feature    1534..1566
                     /label=lox2722
                     /note="color: #faffaf"
     misc_feature    1618..1650
                     /label=loxP
                     /note="color: #ffcd9e"
     CDS             complement(1660..2373)
                     /codon_start=1
                     /label=mRuby3
                     /note="color: #ff0000"
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTD
                     IALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAV
                     AKYSNLGGGMDELYK"
     misc_feature    complement(2383..2760)
                     /label=H2B
                     /note="color: #a6acb3; direction: LEFT"
     CDS             complement(2767..2823)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /note="color: #cc99b2"
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(2839..3033)
                     /codon_start=1
                     /label=Soma-targeting motif
                     /note="color: #339966"
                     /translation="QSQPILNTKEMAPQSKPPEELEMSSMPSPVAPLPARTEGVIDMRS
                     MSSIDSFISCATDFPEATRF"
     CDS             complement(3034..3057)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /note="color: #cc99b2"
                     /translation="DYKDDDDK"
     misc_feature    3058..3072
                     /label=Linker
                     /note="This feature has 2 segments:
                        1: 3058 .. 3070 / #a6acb3
                        2: 3071 .. 3072 / #a6acb3"
     CDS             complement(3073..4047)
                     /codon_start=1
                     /label=Chronos
                     /note="color: #a6acb3"
                     /translation="METAATMTHAFISAVPSAEATIRGLLSAAAVVTPAADAHGETSNA
                     TTAGADHGCFPHINHGTELQHKIAVGLQWFTVIVAIVQLIFYGWHSFKATTGWEEVYVC
                     VIELVKCFIELFHDVDSPATVYQTNGGAVAWLRYSEWLLTCPVILIHLSNLTGLHEEYS
                     KRTMTILVTDIGNIVWGITAAFTKGPLKILFFMIGLFYGVTCFFQIAKVYIESYHTLPK
                     GVCRKICKIMAYVFFCSWLMFPVMFIAGHEGLGLITPYTSGIGHLILDLIAKNTWGFLG
                     HHLRVKIHEHILIHGDIRKTTTINVAGENMEIETFVDEEEEGGV"
     misc_feature    3802..3819
                     /label=seq primer 3
                     /note="color: #a6acb3"
     primer_bind     complement(4031..4067)
                     /label=222_R
                     /note="color: black; sequence: 
                     tacgaagttatgctagcagtatggaaacagccgccac"
     primer_bind     complement(4033..4067)
                     /label=S273A_AAV_R
                     /note="color: black; sequence: 
                     tacgaagttatgctagcagtatggaaacagccgcc"
     misc_feature    4058..4089
                     /label=lox2722(1)
                     /note="color: #fffab7"
     misc_feature    4141..4173
                     /label=loxP(1)
                     /note="color: #ff604e"
     misc_feature    4187..4192
                     /label=HindIII
                     /note="color: #fffba2"
     misc_feature    4193..4797
                     /label=WPRE
                     /note="color: #47ff46"
     misc_feature    4836..5048
                     /label=SV40 polyA
                     /note="color: #996633"
     misc_feature    5119..5249
                     /label=3' ITR
                     /note="color: #00ffff"
     misc_feature    5431..5748
                     /label=f1 origin
                     /note="color: #ff8000"
     misc_feature    6012..6871
                     /label=Amp
                     /note="color: #00ffff"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
       61 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      121 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      181 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      241 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      301 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccagattt
      361 aattaaggcc ttaattaggc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc
      421 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
      481 agtggccaac tccatcacta ggggttcctt gtagttaatg attaacccgc catgctactt
      541 atctacgtag ccatgctcta ggaagatctg taccattgac gtcaataatg acgtatgttc
      601 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      661 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      721 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      781 cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg agccccacgt
      841 tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat ttatttattt
      901 tttaattatt ttgtgcagcg atgggggcgg gggggggggg ggggcgcgcg ccaggcgggg
      961 cggggcgggg cgaggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga
     1021 gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa
     1081 agcgaagcgc gcggcgggcg ggagtcgctg cgacgctgcc ttcgccccgt gccccgctcc
     1141 gccgccgcct cgcgccgccc gccccggctc tgactgaccg cgttactccc acaggtgagc
     1201 gggcgggacg gcccttctcc tccgggctgt aattagcgct tggtttaatg acggcttgtt
     1261 tcttttctgt ggctgcgtga aagccttgag gggctccggg agggcccttt gtgcgggggg
     1321 agcggctcgg ggctgtccgc ggggggacgg ctgccttcgg gggggacggg gcagggcggg
     1381 gttcggcttc tggcgtgtga ccggcggctc tagagcctct gctaaccatg ttcatgcctt
     1441 cttctttttc ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg
     1501 caaagaattg gatcctctag agtcgactcc ggaataactt cgtataggat actttatacg
     1561 aagttatgca gaatggtagc tggattgtag ctgctattag caatatgaaa cctcttaata
     1621 acttcgtata gcatacatta tacgaagtta tggcgcgcct cacttgtaca gctcgtccat
     1681 gccaccacca aggttgctgt atttggcaac tgccacttct ctttgcacta cgtaggtttc
     1741 attgtcactc tcctcgatcc tttccaggcg gtgatcaacg gcatggacac cgggcatctt
     1801 gatgttcccg acggtctttt ttgacctgta agttgtcacg aagttgcagt gcagatggcc
     1861 accaccatca actttcagtg cgatgtcagt gtatcctctc agaccaccat ctgctggata
     1921 catcatctct gtattaggct cccaaccctt ggtcttcttc tgcatcacgg gaccattgga
     1981 gggaaagttt acccctctga ccttgacgtt gtagacgagc tcgccatcct caaggctggt
     2041 gtcctgcgtg acggtgacga ctccaccatc ttcgtatctc gtaactcttt cccaagtaaa
     2101 accctcagga aaggactgtt taaagaaatc agggatgtcg gccgggtact tgataaaggt
     2161 acggctgcca tacatgaacg acgtggcaag aatgtcaaag gcaaatggca ggggtcctcc
     2221 ctcgatgact ttgatcctca tggtttgcac tccctcgtac ggtctgcctt ctccttcacc
     2281 tgtgcatttg aattggtggc cgttgaccga accttccatg accaccttca tacgcatatt
     2341 ttccttgatc agctcttcgc ccttagacac catctcgagt gccttagcgc tggtgtactt
     2401 ggtgatggcc ttagtaccct cggacacggc gtgcttggcc aactccccag gcagcagcag
     2461 gcgcacggcc gtctggatct ccctggaggt gatggtcgag cgcttgttgt aatgcgccag
     2521 gcgggaagcc tcacctgcga tgcgctcgaa aatgtcgttc acaaacgaat tcatgatgcc
     2581 catggccttg gacgaaatgc cggtgtcagg gtggacctgc ttcagaacct tgtacacata
     2641 gatggaatag ctctccttgc ggctgcgctt gcgcttcttg ccgcctttct tctgcgcctt
     2701 agtcaccgcc ttcttggagc cctttttcgg ggcgggagca gacttcgctg gctctggcat
     2761 actagtaggt ccagggttct cctccacgtc tccagcctgc ttcagcaggc tgaagttagt
     2821 agctccgctt cctgccgcga atctggtggc ttcagggaag tccgtggcac agctgatgaa
     2881 gctgtcaatg ctggacatgc tccgcatgtc gatgacgccc tccgtgcgtg cgggcagagg
     2941 ggccacgggg ctgggcatgc tgctcatctc cagctcctct ggaggcttgc tctgcggggc
     3001 catctccttg gtgttgagga tgggctggga ctgcttatcg tcatcgtctt tgtaatctac
     3061 cggtgcggcc gccactcctc cctcctcctc ctcgtcgacg aaggtctcga tctccatgtt
     3121 ctcgccggcc acgttgatgg tggttgtctt ccggatgtcg ccgtggatca ggatgtgctc
     3181 gtggatcttc actctcaggt ggtggcccag gaagccccaa gtgttcttgg cgatcagatc
     3241 caggatcagg tggccgattc cgctggtgta aggtgtgatc aggcccagtc cctcgtgtcc
     3301 ggcgatgaac atcacgggga acatcagcca agagcagaag aagacgtagg ccatgatctt
     3361 gcaaatcttc cggcagacgc ctttgggcag ggtgtggtag ctctcgatat acaccttggc
     3421 gatctggaag aagcaagtca cgccgtagaa caggccgatc atgaagaaca ggatcttcag
     3481 ggggcccttt gtaaaggcgg ctgtgatccc ccacacgatg ttgccgatgt cggtcaccag
     3541 gatggtcatg gtccgcttgc tgtactcttc gtgcagtccg gtcaggttgc tcaggtggat
     3601 caggatcacg gggcaagtca ggagccactc gctgtaccgc agccaagcca cggctcctcc
     3661 gttggtctgg tacactgtgg ctgggctgtc gacatcgtgg aacagctcga tgaagcactt
     3721 gacgagctcg atcacgcaga cgtagacctc ctcccagccg gttgtggcct tgaagctgtg
     3781 ccaaccgtag aagatgagct gcacgatagc cacgatcacg gtgaaccact ggagtcccac
     3841 tgcgatcttg tgctgcagct cggttccgtg gttgatgtgg gggaagcaac cgtgatcggc
     3901 tccggctgtt gtggcgttag aggtttctcc gtgagcgtct gctgctggtg tcaccactgc
     3961 tgcggcgctc agcaggcctc taattgtggc ttcggcgcta ggcacggctg agataaaggc
     4021 gtgggtcatt gtggcggctg tttccatact gctagcataa cttcgtataa agtatcctat
     4081 acgaagttat ttgccttaac ccagaaatta tcactgttat tctttagaat ggtgcaaaga
     4141 ataacttcgt ataatgtatg ctatacgaag ttatgaattc gatatcaagc ttatcgataa
     4201 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     4261 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     4321 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     4381 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     4441 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     4501 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     4561 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     4621 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     4681 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     4741 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg
     4801 acctcgaccc gggcggccgc ttcgagcaga catgataaga tacattgatg agtttggaca
     4861 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     4921 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     4981 tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa acctctacaa
     5041 atgtggtaaa atcgataagg atcttcctag agcatggcta cgtagataag tagcatggcg
     5101 ggttaatcat taactacaag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     5161 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     5221 cggcctcagt gagcgagcga gcgcgcagcc ttaattaacc taattcactg gccgtcgttt
     5281 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     5341 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     5401 tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     5461 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     5521 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     5581 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     5641 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     5701 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     5761 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     5821 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     5881 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     5941 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     6001 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     6061 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     6121 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     6181 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     6241 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     6301 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     6361 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     6421 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     6481 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     6541 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     6601 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     6661 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     6721 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     6781 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6841 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     6901 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6961 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     7021 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     7081 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     7141 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     7201 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     7261 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     7321 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     7381 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     7441 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     7501 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     7561 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     7621 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7681 tgctcacatg
//
