LOCUS       Exported                7723 bp ds-DNA     circular SYN 26-FEB-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7723)
  AUTHORS   Adesnik Lab
  TITLE     Direct Submission
  JOURNAL   Exported Oct 1, 2024 from SnapGene 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7723
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     misc_feature    378..509
                     /label=5'ITR
     promoter        823..1100
                     /label=chicken beta-actin promoter
     misc_feature    1511..1515
                     /label=BamHI
     misc_feature    1534..1566
                     /label=lox2722
     misc_feature    1618..1650
                     /label=loxP
     CDS             complement(1660..2373)
                     /codon_start=1
                     /label=mRuby3
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTD
                     IALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAV
                     AKYSNLGGGMDELYK"
     misc_feature    complement(2383..2760)
                     /label=H2B
     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."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(2857..3051)
                     /codon_start=1
                     /label=ST
                     /translation="QSQPILNTKEMAPQSKPPEELEMSSMPSPVAPLPARTEGVIDMRS
                     MSSIDSFISCATDFPEATRF"
     CDS             complement(3052..3075)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     3066..3097
                     /label=314_F
     misc_feature    3076..3082
                     /label=Linker
     misc_feature    complement(3085..3144)
                     /label=Surface expression
     CDS             complement(3154..4080)
                     /codon_start=1
                     /label=ChRmine
                     /translation="MAHAPGTDQMFYVGTMDGWYLDTKLNSVAIGAHWSCFIVLTITTF
                     YLGYESWTSRGPSKRTSFYAGYQEEQNLALFVNFFAMLSYFGKIVADTLGHNFGDVGPF
                     IIGFGNYRYADYMLTCPMLVYDLLYQLRAPYRVSCSAIIFAILMSGVLAEFYAEGDPRL
                     RNGAYAWYGFGCFWFIFAYSIVMSIVAKQYSRLAQLAQDTGAEHSLHVLKFAVFTFSML
                     WILFPLVWAICPRGFGWIDDNWTEVAHCVCDIVAKSCYGFALARFRKTYDEELFRLLEQ
                     LGHDEDEFQKLELDMRLSSNGERLRRLS"
     primer_bind     complement(4068..4100)
                     /label=314_R
     misc_feature    4091..4122
                     /label=lox2722(1)
     misc_feature    4174..4206
                     /label=loxP(1)
     misc_feature    4220..4225
                     /label=HindIII
     misc_feature    4226..4830
                     /label=WPRE
     misc_feature    4869..5081
                     /label=SV40 polyA
     misc_feature    5152..5282
                     /label=3' ITR
     misc_feature    5464..5781
                     /label=f1 origin
     misc_feature    6045..6904
                     /label=Amp
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 cctgccacct cgttctcgta gcagaagaat ctggtggctt cagggaagtc
     2881 cgtggcacag ctgatgaagc tgtcaatgct ggacatgctc cgcatgtcga tgacgccctc
     2941 cgtgcgtgcg ggcagagggg ccacggggct gggcatgctg ctcatctcca gctcctctgg
     3001 aggcttgctc tgcggggcca tctccttggt gttgaggatg ggctgggact gcttatcgtc
     3061 atcgtctttg taatctaccg gtaccacgtt gatgtcgatc tggtccaggg ggatgtactc
     3121 gccctcgctg gtgatcctgc tcttggcggc cgcagacagt ctccgcaggc gctcgccatt
     3181 gctgctcagc ctcatatcca gctccagctt ctggaactca tcctcgtcgt gtcccagctg
     3241 ctccagcagc cgaaacagct cctcatcata ggtctttctg aaccgggcca gggcaaagcc
     3301 gtaacaggac ttggccacga tgtcgcacac gcagtgggcc acctctgtcc agttatcgtc
     3361 gatccagccg aagcctctag ggcagatggc ccacaccagg ggaaacagaa tccacagcat
     3421 ggagaaggta aacacggcga acttcagcac gtgcaggctg tgctcggcgc ctgtatcctg
     3481 ggccagctgg gccagccggc tatactgctt ggccacgatg ctcatcacga tagagtaggc
     3541 gaagataaac cagaaacagc caaagccata ccaggcgtag gcgccattcc tcagcctagg
     3601 gtcgccctct gcgtagaact cggccagcac gccagacatc aggatggcaa agatgatggc
     3661 gctacaagac acgcgatagg gggccctcag ctgatacagc agatcgtaca ccagcattgg
     3721 gcaggtcagc atgtagtcgg catacctgta attgccgaag ccgatgatga aggggcccac
     3781 atcgccgaag ttgtggccca gtgtgtcggc cacgatcttg ccgaagtagc tcagcatggc
     3841 aaagaaattc acaaacaggg ccaggttctg ctcctcctga tagccggcgt aaaaggaggt
     3901 tctcttgcta gggccccgag atgtccagga ctcatagccc aggtagaatg tggtgattgt
     3961 cagcacgata aagcaagacc agtgggcgcc gatggccacg gagttcagct tggtatccag
     4021 ataccagccg tccattgtgc ccacgtagaa catctggtcg gtgcctggtg cgtgtgccat
     4081 ggtgctagca taacttcgta taaagtatcc tatacgaagt tatttgcctt aacccagaaa
     4141 ttatcactgt tattctttag aatggtgcaa agaataactt cgtataatgt atgctatacg
     4201 aagttatgaa ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga
     4261 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     4321 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     4381 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     4441 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     4501 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     4561 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     4621 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     4681 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     4741 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     4801 cctttgggcc gcctccccgc atcgataccg tcgacctcga cccgggcggc cgcttcgagc
     4861 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     4921 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     4981 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg
     5041 ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatcttcc
     5101 tagagcatgg ctacgtagat aagtagcatg gcgggttaat cattaactac aaggaacccc
     5161 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     5221 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     5281 gccttaatta acctaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     5341 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     5401 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg
     5461 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     5521 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     5581 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     5641 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     5701 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     5761 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     5821 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     5881 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     5941 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     6001 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     6061 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     6121 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     6181 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     6241 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     6301 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     6361 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     6421 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     6481 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     6541 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     6601 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     6661 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     6721 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     6781 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     6841 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     6901 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     6961 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     7021 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     7081 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     7141 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     7201 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     7261 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     7321 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     7381 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     7441 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     7501 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     7561 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     7621 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     7681 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atg
//
