LOCUS       40924_32973        5681 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pN16-DsRed-IRES-GFP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5681)
  AUTHORS   Li I-C., Chiu C-Y., Wu C-L., Chi J-Y., Jian S-R., Wang S-W., Chang 
            CL.
  TITLE     A dual-fluorescent reporter system facilitates identification of 
            thiol compounds that suppress oxidative frameshift mutations
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5681)
  AUTHORS   Jian S-R., Chang CL.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-MAY-2012) Institute of Molecular Medicine, National 
            Cheng Kung University, 1 Ta-Hsueh Road, Tainan, Taiwan 70101, 
            Republic of China
REFERENCE   3  (bases 1 to 5681)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5681)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (22-MAY-2012) Institute of Molecular Medicine, National Cheng Kung 
            University, 1 Ta-Hsueh Road, Tainan, Taiwan 70101, Republic of 
            China"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5681
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        67..370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        371..574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        620..638
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    660..675
                     /label=N16 random sequence
                     /note="N16 random sequence"
     CDS             689..1366
                     /codon_start=1
                     /label=DsRed1
                     /note="wild-type DsRed"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLFL"
     CDS             1418..1489
                     /codon_start=1
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags"
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     misc_feature    1525..2108
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             2109..2825
                     /codon_start=1
                     /label=hrGFP
                     /note="humanized Renilla green fluorescent protein"
                     /translation="MVSKQILKNTGLQEIMSFKVNLEGVVNNHVFTMEGCGKGNILFGN
                     QLVQIRVTKGAPLPFAFDILSPAFQYGNRTFTKYPEDISDFFIQSFPAGFVYERTLRYE
                     DGGLVEIRSDINLIEEMFVYRVEYKGRNFPNDGPVMKKTITGLQPSFEVVYMNDGVLVG
                     QVILVYRLNSGKFYSCHMRTLMKSKGVVKDFPEYHFIQHRLEKTYVEDGGFVEQHETAI
                     AQLTSLGKPLGSLHEWV"
     promoter        complement(2859..2877)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3151..3272
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3279..3734)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3761..3863
                     /label=AmpR promoter
     protein_bind    3880..3913
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(3958..4815)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4985..5573
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
       61 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      541 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      601 gcgattacgc caagctcgaa attaaccctc actaaaggga acaaaagctg gagctcatgg
      661 atatcattac tagtaaccgg tcgccaccat ggtgcgctcc tccaagaacg tcatcaagga
      721 gttcatgcgc ttcaaggtgc gcatggaggg caccgtgaac ggccacgagt tcgagatcga
      781 gggcgagggc gagggccgcc cctacgaggg ccacaacacc gtgaagctga aggtgaccaa
      841 gggcggcccc ctgcccttcg cctgggacat cctgtccccc cagttccagt acggctccaa
      901 ggtgtacgtg aagcaccccg ccgacatccc cgactacaag aagctgtcct tccccgaggg
      961 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga
     1021 ctcctccctg caggacggct gcttcatcta caaggtgaag ttcatcggcg tgaacttccc
     1081 ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcca ccgagcgcct
     1141 gtacccccgc gacggcgtgc tgaagggcga gatccacaag gccctgaagc tgaaggacgg
     1201 cggccactac ctggtggagt tcaagtccat ctacatggcc aagaagcccg tgcagctgcc
     1261 cggctactac tacgtggact ccaagctgga catcacctcc cacaacgagg actacaccat
     1321 cgtggagcag tacgagcgca ccgagggccg ccaccacctg ttcctgtagc ggccgctcta
     1381 gcccgggcgg atccgaattc gcatgcgtcg actcgaggac tacaaggatg acgatgacaa
     1441 ggattacaaa gacgacgatg ataaggacta taaggatgat gacgacaaat aatagcaatt
     1501 cctcgacgac tgcatagggt tacccccctc tccctccccc ccccctaacg ttactggccg
     1561 aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc
     1621 gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag
     1681 gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt
     1741 tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa
     1801 ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc
     1861 aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg
     1921 gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat
     1981 gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa
     2041 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatgg
     2101 ccacaaccat ggtgagcaag cagatcctga agaacaccgg cctgcaggag atcatgagct
     2161 tcaaggtgaa cctggagggc gtggtgaaca accacgtgtt caccatggag ggctgcggca
     2221 agggcaacat cctgttcggc aaccagctgg tgcagatccg cgtgaccaag ggcgcccccc
     2281 tgcccttcgc cttcgacatc ctgagccccg ccttccagta cggcaaccgc accttcacca
     2341 agtaccccga ggacatcagc gacttcttca tccagagctt ccccgccggc ttcgtgtacg
     2401 agcgcaccct gcgctacgag gacggcggcc tggtggagat ccgcagcgac atcaacctga
     2461 tcgaggagat gttcgtgtac cgcgtggagt acaagggccg caacttcccc aacgacggcc
     2521 ccgtgatgaa gaagaccatc accggcctgc agcccagctt cgaggtggtg tacatgaacg
     2581 acggcgtgct ggtgggccag gtgatcctgg tgtaccgcct gaacagcggc aagttctaca
     2641 gctgccacat gcgcaccctg atgaagagca agggcgtggt gaaggacttc cccgagtacc
     2701 acttcatcca gcaccgcctg gagaagacct acgtggagga cggcggcttc gtggagcagc
     2761 acgagaccgc catcgcccag ctgaccagcc tgggcaagcc cctgggcagc ctgcacgagt
     2821 gggtgtaata gggtaccagg taagtgtacc caattcgccc tatagtgagt cgtattacaa
     2881 ttcactcgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga gatccaattt
     2941 ttaagtgtat aatgtgttaa actactgatt ctaattgttt gtgtatttta gattcacagt
     3001 cccaaggctc atttcaggcc cctcagtcct cacagtctgt tcatgatcat aatcagccat
     3061 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     3121 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     3181 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3241 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt
     3301 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     3361 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     3421 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     3481 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     3541 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     3601 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     3661 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     3721 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3781 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3841 tcaataatat tgaaaaagga agaatcctga ggccgggcca taacttcgta taatgtatgc
     3901 tatacgaagt tatccatggg ccccccctcg acatgagtaa acttggtctg acagttacca
     3961 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4021 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4081 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4141 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4201 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4261 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4321 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4381 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4441 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4501 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4561 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4621 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4681 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4741 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4801 atgttgaata ctcatactct tcctcaggtt actcatatat actttagatt gatttaaaac
     4861 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4921 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4981 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5041 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5101 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     5161 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5221 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5281 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5341 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5401 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5461 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5521 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5581 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     5641 ctgcgttatc ccctgattct gtggataacc gtattaccgc c
//
