LOCUS       sequence-141472-        5740 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCS4+-NLS-EGFP-F2A-mCherry-CAAX.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5740)
  AUTHORS   Kim JH, Lee SR, Li LH, Park HJ, Park JH, Lee KY, Kim MK, Shin BA, 
            Choi SY
  TITLE     High Cleavage Efficiency of a 2A Peptide Derived from Porcine 
            Teschovirus-1 in Human Cell Lines, Zebrafish and Mice.
  JOURNAL   PLoS One. 2011  . 6(4):e18556.
  PUBMED    21602908
REFERENCE   2  (bases 1 to 5740)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One. 
            2011  . 6(4):e18556."
COMMENT     Sequence Label: pSYC-104
FEATURES             Location/Qualifiers
     source          1..5740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..987
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1022..1040
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1022..1039
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1088..1108
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1154..1156,1157..1159,1160..1870)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1154..1156,1157..1159,1160..1870)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1199..1220)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1460..1479)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1807..1828
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1898..1963
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     CDS             1973..2680
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2118..2136)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2386..2405
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2580..2603
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2752..2771)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2753..2771)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2776..2910
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2800..2819)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2854..2873
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3023..3043)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3023..3041)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3062..3078)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3062..3078)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3075..3097)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3086..3102
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3110..3116,3117..3134,3135..3140))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3155..3176
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3293..3310)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3464..4052)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3544..3563)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4223..5014,5015..5083))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4846..4865
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5084..5188)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5214..5669)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5351..5372)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5563..5582
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tcgaccatag ccaattcaat atggcgtata tggactcatg ccaattcaat atggtggatc
       61 tggacctgtg ccaattcaat atggcgtata tggactcgtg ccaattcaat atggtggatc
      121 tggaccccag ccaattcaat atggcggact tggcaccatg ccaattcaat atggcggact
      181 tggcactgtg ccaactgggg aggggtctac ttggcacggt gccaagtttg aggaggggtc
      241 ttggccctgt gccaagtccg ccatattgaa ttggcatggt gccaataatg gcggccatat
      301 tggctatatg ccaggatcaa tatataggca atatccaata tggccctatg ccaatatggc
      361 tattggccag gttcaatact atgtattggc cctatgccat atagtattcc atatatgggt
      421 tttcctattg acgtagatag cccctcccaa tgggcggtcc catataccat atatggggct
      481 tcctaatacc gcccatagcc actcccccat tgacgtcaat ggtctctata tatggtcttt
      541 cctattgacg tcatatgggc ggtcctattg acgtatatgg cgcctccccc attgacgtca
      601 attacggtaa atggcccgcc tggctcaatg cccattgacg tcaataggac cacccaccat
      661 tgacgtcaat gggatggctc attgcccatt catatccgtt ctcacgcccc ctattgacgt
      721 caatgacggt aaatggccca cttggcagta catcaatatc tattaatagt aacttggcaa
      781 gtacattact attggaagga cgccagggta cattggcagt actcccattg acgtcaatgg
      841 cggtaaatgg cccgcgatgg ctgccaagta catccccatt gacgtcaatg gggaggggca
      901 atgacgcaaa tgggcgttcc attgacgtaa atgggcggta ggcgtgccta atgggaggtc
      961 tatataagca atgctcgttt agggaaccgc cattctgcct ggggacgtcg gagcaagctt
     1021 gatttaggtg acactataga atacaagcta cttgttcttt ttgcaggatc ccatcgatac
     1081 catggctcca aagaagaagc gtaaggtaaa tcgatcgaat tcaaggcctc tcgagcctct
     1141 agaaggtggc ggaatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     1201 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     1261 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     1321 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     1381 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     1441 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     1501 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     1561 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     1621 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     1681 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     1741 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     1801 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     1861 gctgtacaag cttaaggagc tcgcatgcgg aagcggagtg aaacagactt tgaattttga
     1921 ccttctcaag ttggcgggag acgtggagtc caaccctgga cctagatcta ccatggtgag
     1981 caagggcgag gaggacaaca tggccatcat caaggagttc atgcgcttca aggtgcacat
     2041 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
     2101 cgagggcacc cagaccgcca agctgaaggt gaccaagggc ggccccctgc ccttcgcctg
     2161 ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga
     2221 catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     2281 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt
     2341 catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa
     2401 gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa
     2461 gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg ccgaggtcaa
     2521 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa
     2581 gctggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg agcgcgccga
     2641 gggccgccac tccaccggcg gcatggacga gctgtacaag aagctgaacc ctcctgatga
     2701 gagtggcccc ggctgcatga gctgcaagtg tgtgctctcc tgagctagcc tccctatagt
     2761 gagtcgtatt acgtagatcc agacatgata agatacattg atgagtttgg acaaaccaca
     2821 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     2881 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     2941 caggttcagg gggaggtgtg ggaggttttt taattcgcgg ccgcggcgcc aatgcattgg
     3001 gcccggtacc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
     3061 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     3121 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     3181 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     3241 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3301 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3361 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3421 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3481 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3541 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3601 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3661 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3721 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3781 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3841 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3901 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3961 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4021 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4081 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4141 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4201 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4261 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4321 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4381 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4441 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4501 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4561 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4621 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4681 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4741 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4801 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     4861 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     4921 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     4981 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5041 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5101 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5161 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt
     5221 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     5281 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     5341 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     5401 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     5461 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     5521 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     5581 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     5641 cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg
     5701 ggaagggcga tcggtgcggg cctcttcgct attacgccag
//
