LOCUS       Exported                5698 bp ds-DNA     circular SYN 02-NOV-2018
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5698)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Aug 13, 2020 from SnapGene 4.3.11
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5698
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     150..172
                     /label=012 SV40-F
                     /note="color: black; sequence: 
                     GTGCTAGCCCGGGCTCGATGCATCTCAATTAGTCAGCAAC"
     primer_bind     222..244
                     /label=012 SV40-F
                     /note="color: black; sequence: 
                     GTGCTAGCCCGGGCTCGATGCATCTCAATTAGTCAGCAAC"
     primer_bind     complement(399..419)
                     /label=013 SV40-R
                     /note="color: black; sequence: 
                     AAGCTTACTTAGATCGCATTTGCAAAAGCCTAGGCCTCC"
     primer_bind     607..624
                     /label=SI003/psiC-5UTR-seq
                     /note="psiCHECK2に入れたinsertを読むのに使う"
                     /note="color: black; sequence: CCTTTCTCTCCACAGGTG"
     misc_binding    666..684
                     /note="color: #ff6666"
     primer_bind     666..684
                     /label=T7 primer
                     /note="color: black; sequence: TAATACGACTCACTATAGG"
     primer_bind     666..684
                     /label=T7 promoter
                     /note="color: black; sequence: tgacTAATACGACTCACTATAGG"
     5'UTR           685..705
                     /label=EIF2S3 5UTR
                     /note="color: #a6acb3; direction: RIGHT"
     CDS             706..1638
                     /codon_start=1
                     /label=hRluc-Renilla
                     /note="color: #4444ff"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     primer_bind     706..726
                     /label=006 hRluc ATG-F
                     /note="psiCHECK2のhRlucの頭のprimerです。004&006で増やしたhRlucの一部をG.
                     Aで入れ込むのに使用"
                     /note="color: black; sequence: ATGGCTTCCAAGGTGTACGAC"
     primer_bind     complement(841..858)
                     /label=SI004/hRluc-toe
                     /note="psiCHECK2に入れたinsertを後ろから読むprimer"
                     /note="color: black; sequence: ATGCAGAAAAATCACGGC"
     primer_bind     1088..1106
                     /label=SI044
                     /note="color: black; sequence: ACTACTCCTACGAGCACCA"
     primer_bind     complement(1168..1209)
                     /label=004 hRluc EcoRV-2R
                     /note="psiCHECK2ベクターをEcoRVで切った時にG.Aでつながるprimerです。"
                     /note="color: black; sequence: 
                     GCTCTTGATCAGGGCGATATCCTCCTCGATGTCAGGCCACTC"
     primer_bind     1190..1209
                     /label=121 hRluc seq-1F
                     /note="color: black; sequence: ATATCGCCCTGATCAAGAGC"
     primer_bind     1569..1587
                     /label=SI045
                     /note="color: black; sequence: GGAGGACGCTCCAGATGAA"
     primer_bind     1618..1657
                     /label=SI082
                     /note="color: black; sequence: 
                     CGCGTGCTGAAGAACGAGCAGGGAAGCGGAGCTACTAACT"
     primer_bind     complement(1618..1638)
                     /label=SI035
                     /note="color: black; sequence: CTGCTCGTTCTTCAGCACGCG"
     CDS             1639..1704
                     /codon_start=1
                     /label=2A Porcine teschovirus-1 (PTV1)
                     /note="color: #663300"
                     /translation="GSGATNFSLLKQAGDVEENPGP"
     primer_bind     1639..1657
                     /label=SI093
                     /note="color: black; sequence: GGAAGCGGAGCTACTAACT"
     primer_bind     complement(1688..1705)
                     /label=SI083
                     /note="color: black; sequence: 
                     CTTCTTAATGTTCTTAGCATCGGCAGGTCCAGGGTTCTCCT"
     misc_feature    1699..1704
                     /label=cleavage site
                     /note="color: #a6acb3"
     CDS             1705..1770
                     /codon_start=1
                     /label=3xFLAG
                     /note="color: #dd0000"
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1748..1770)
                     /label=SI092
                     /note="color: black; sequence: CTTGTCATCGTCATCCTTGTAAT"
     primer_bind     1756..1786
                     /label=SI094
                     /note="color: black; sequence: 
                     GATGACGATGACAAGGCCTGGAGGAGCTCCC"
     CDS             1771..1878
                     /codon_start=1
                     /label=Reporter Seq
                     /note="color: #993366"
                     /translation="AWRSSQRSTQKDPVPYQPPFLCQWGRHQPSWKPLMN"
     primer_bind     1771..1840
                     /label=SI110
                     /note="color: black; sequence: 
                     GCCTGGAGGAGCTCCCAGAGGTCTACCCAGAAGGACCCAGTTCCTTACCAGCCTCCCTT
                     TCTCTGTCAGT"
     primer_bind     complement(1824..1893)
                     /label=SI111
                     /note="color: black; sequence: 
                     GGTCAAAACGAATGAGTTCATTAATGGCTTCCAGCTTGGCTGATGACGTCCCCACTGAC
                     AGAGAAAGGGA"
     primer_bind     complement(1857..1908)
                     /label=SI095
                     /note="color: black; sequence: 
                     AGTAGCTCCGCTTCCGGTCAAAACGAATGAGTTCATTAATGGCTTCCAGCTT"
     CDS             1876..1878
                     /codon_start=1
                     /label=Pause site
                     /note="color: #993366"
                     /translation="N"
     CDS             1879..1893
                     /codon_start=1
                     /label=Mouse XBP1u seq followed pause
                     /note="color: #993366"
                     /translation="SFVLT"
     CDS             1894..1959
                     /codon_start=1
                     /label=2A Porcine teschovirus-1 (PTV1)
                     /note="color: #663300"
                     /translation="GSGATNFSLLKQAGDVEENPGP"
     primer_bind     1894..1912
                     /label=SI093
                     /note="color: black; sequence: GGAAGCGGAGCTACTAACT"
     primer_bind     1894..1912
                     /label=SI082
                     /note="color: black; sequence: 
                     CGCGTGCTGAAGAACGAGCAGGGAAGCGGAGCTACTAACT"
     primer_bind     complement(1943..1983)
                     /label=SI083
                     /note="color: black; sequence: 
                     CTTCTTAATGTTCTTAGCATCGGCAGGTCCAGGGTTCTCCT"
     misc_feature    1955..1959
                     /label=cleavage site
                     /note="color: #a6acb3"
     CDS             1960..3609
                     /codon_start=1
                     /label=hluc+ FL
                     /note="color: #ff8000"
                     /translation="ADAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDAH
                     IEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAPA
                     NDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSM
                     YTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHAR
                     DPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYKI
                     QSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYG
                     LTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMSG
                     YVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESILL
                     QHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVF
                     VDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     1960..1983
                     /label=SI081
                     /note="color: black; sequence: GCCGATGCTAAGAACATTAAGAAG"
     primer_bind     1960..1980
                     /label=110 HCF_IRES_FL hluc-F
                     /note="color: black; sequence: 
                     AAAAACCAAACGTAACATGGCCGATGCTAAGAACATTAAG"
     primer_bind     1960..1979
                     /label=SI034
                     /note="color: black; sequence: ATGGCCGATGCTAAGAACATTAA"
     primer_bind     complement(1962..1983)
                     /label=CTTCTTAATGTTCTTAGCATCG
                     /note="color: black; sequence: CTTCTTAATGTTCTTAGCATCG"
     primer_bind     2369..2386
                     /label=SI046
                     /note="color: black; sequence: ACGTGCAGAAGAAGCTGC"
     primer_bind     2528..2549
                     /label=112 Ffluc-1F
                     /note="color: black; sequence: CCATTGCCCTGATCATGAACAG"
     primer_bind     2849..2868
                     /label=SI047
                     /note="color: black; sequence: GCACCCTGATCGACAAGTAC"
     primer_bind     3330..3348
                     /label=SI048
                     /note="color: black; sequence: GCTGCAGCACCCTAACATT"
     primer_bind     complement(3637..3661)
                     /label=Fluc R　SI016
                     /note="color: black; sequence: TGTATCTTATCATGTCTGCTCGAAG"
     terminator      3644..3865
                     /label=SV40 late poly(A)
                     /note="color: #ffff00"
ORIGIN
        1 agatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct
       61 tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
      121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
      181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
      241 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
      301 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
      361 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
      421 agcttgattc ttctgacaca acagtctcga acttaagctg cagaagttgg tcgtgaggca
      481 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
      541 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
      601 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga
      661 gtacttaata cgactcacta taggtttcct tcctcttttg gcaacatggc ttccaaggtg
      721 tacgaccccg agcaacgcaa acgcatgatc actgggcctc agtggtgggc tcgctgcaag
      781 caaatgaacg tgctggactc cttcatcaac tactatgatt ccgagaagca cgccgagaac
      841 gccgtgattt ttctgcatgg taacgctgcc tccagctacc tgtggaggca cgtcgtgcct
      901 cacatcgagc ccgtggctag atgcatcatc cctgatctga tcggaatggg taagtccggc
      961 aagagcggga atggctcata tcgcctcctg gatcactaca agtacctcac cgcttggttc
     1021 gagctgctga accttccaaa gaaaatcatc tttgtgggcc acgactgggg ggcttgtctg
     1081 gcctttcact actcctacga gcaccaagac aagatcaagg ccatcgtcca tgctgagagt
     1141 gtcgtggacg tgatcgagtc ctgggacgag tggcctgaca tcgaggagga tatcgccctg
     1201 atcaagagcg aagagggcga gaaaatggtg cttgagaata acttcttcgt cgagaccatg
     1261 ctcccaagca agatcatgcg gaaactggag cctgaggagt tcgctgccta cctggagcca
     1321 ttcaaggaga agggcgaggt tagacggcct accctctcct ggcctcgcga gatccctctc
     1381 gttaagggag gcaagcccga cgtcgtccag attgtccgca actacaacgc ctaccttcgg
     1441 gccagcgacg atctgcctaa gatgttcatc gagtccgacc ctgggttctt ttccaacgct
     1501 attgtcgagg gagctaagaa gttccctaac accgagttcg tgaaggtgaa gggcctccac
     1561 ttcagccagg aggacgctcc agatgaaatg ggtaagtaca tcaagagctt cgtggagcgc
     1621 gtgctgaaga acgagcaggg aagcggagct actaacttca gcctgctgaa gcaggctgga
     1681 gacgtggagg agaaccctgg acctgactac aaagaccatg acggtgatta taaagatcat
     1741 gacatcgatt acaaggatga cgatgacaag gcctggagga gctcccagag gtctacccag
     1801 aaggacccag ttccttacca gcctcccttt ctctgtcagt ggggacgtca tcagccaagc
     1861 tggaagccat taatgaactc attcgttttg accggaagcg gagctactaa cttcagcctg
     1921 ctgaagcagg ctggagacgt ggaggagaac cctggacctg ccgatgctaa gaacattaag
     1981 aagggccctg ctcccttcta ccctctggag gatggcaccg ctggcgagca gctgcacaag
     2041 gccatgaaga ggtatgccct ggtgcctggc accattgcct tcaccgatgc ccacattgag
     2101 gtggacatca cctatgccga gtacttcgag atgtctgtgc gcctggccga ggccatgaag
     2161 aggtacggcc tgaacaccaa ccaccgcatc gtggtgtgct ctgagaactc tctgcagttc
     2221 ttcatgccag tgctgggcgc cctgttcatc ggagtggccg tggcccctgc taacgacatt
     2281 tacaacgagc gcgagctgct gaacagcatg ggcatttctc agcctaccgt ggtgttcgtg
     2341 tctaagaagg gcctgcagaa gatcctgaac gtgcagaaga agctgcctat catccagaag
     2401 atcatcatca tggactctaa gaccgactac cagggcttcc agagcatgta cacattcgtg
     2461 acatctcatc tgcctcctgg cttcaacgag tacgacttcg tgccagagtc tttcgacagg
     2521 gacaaaacca ttgccctgat catgaacagc tctgggtcta ccggcctgcc taagggcgtg
     2581 gccctgcctc atcgcaccgc ctgtgtgcgc ttctctcacg cccgcgaccc tattttcggc
     2641 aaccagatca tccccgacac cgctattctg agcgtggtgc cattccacca cggcttcggc
     2701 atgttcacca ccctgggcta cctgatttgc ggctttcggg tggtgctgat gtaccgcttc
     2761 gaggaggagc tgttcctgcg cagcctgcaa gactacaaaa ttcagtctgc cctgctggtg
     2821 ccaaccctgt tcagcttctt cgctaagagc accctgatcg acaagtacga cctgtctaac
     2881 ctgcacgaga ttgcctctgg cggcgcccca ctgtctaagg aggtgggcga agccgtggcc
     2941 aagcgctttc atctgccagg catccgccag ggctacggcc tgaccgagac aaccagcgcc
     3001 attctgatta ccccagaggg cgacgacaag cctggcgccg tgggcaaggt ggtgccattc
     3061 ttcgaggcca aggtggtgga cctggacacc ggcaagaccc tgggagtgaa ccagcgcggc
     3121 gagctgtgtg tgcgcggccc tatgattatg tccggctacg tgaataaccc tgaggccaca
     3181 aacgccctga tcgacaagga cggctggctg cactctggcg acattgccta ctgggacgag
     3241 gacgagcact tcttcatcgt ggaccgcctg aagtctctga tcaagtacaa gggctaccag
     3301 gtggccccag ccgagctgga gtctatcctg ctgcagcacc ctaacatttt cgacgccgga
     3361 gtggccggcc tgcccgacga cgatgccggc gagctgcctg ccgccgtcgt cgtgctggaa
     3421 cacggcaaga ccatgaccga gaaggagatc gtggactatg tggccagcca ggtgacaacc
     3481 gccaagaagc tgcgcggcgg agtggtgttc gtggacgagg tgcccaaggg cctgaccggc
     3541 aagctggacg cccgcaagat ccgcgagatc ctgatcaagg ctaagaaagg cggcaagatc
     3601 gccgtgtaat aattctagag tcggggcggc cggccgcttc gagcagacat gataagatac
     3661 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     3721 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     3781 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     3841 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc caggtggcac ttttcgggga
     3901 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3961 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     4021 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     4081 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     4141 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     4201 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     4261 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     4321 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     4381 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     4441 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     4501 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     4561 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4621 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4681 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4741 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4801 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4861 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4921 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4981 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5041 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5101 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5161 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     5221 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5281 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5341 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5401 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5461 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5521 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5581 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5641 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat ggctcgac
//
