LOCUS       sequence-141471-        5731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCS4+-NLS-EGFP-E2A-mCherry-CAAX.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5731)
  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 5731)
  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-103
FEATURES             Location/Qualifiers
     source          1..5731
                     /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..1957
                     /codon_start=1
                     /product="2A peptide from equine rhinitis A virus 
                     polyprotein"
                     /label=E2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="QCTNYALLKLAGDVESNPGP"
     CDS             1967..2674
                     /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(2112..2130)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2380..2399
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2574..2597
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2767..2901
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2791..2810)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2845..2864
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3014..3034)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3014..3032)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3053..3069)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3053..3069)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3066..3088)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3077..3093
                     /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(3101..3107,3108..3125,3126..3131))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3146..3167
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3284..3301)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3455..4043)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3535..3554)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4214..5005,5006..5074))
                     /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     4837..4856
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5075..5179)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5205..5660)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5342..5363)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5554..5573
                     /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 aagcggacag tgtactaatt atgctctctt
     1921 gaaattggct ggagatgttg agagcaaccc tggacctaga tctaccatgg tgagcaaggg
     1981 cgaggaggac aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg
     2041 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     2101 cacccagacc gccaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat
     2161 cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc
     2221 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     2281 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta
     2341 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac
     2401 catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga
     2461 gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgccgagg tcaagaccac
     2521 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagctgga
     2581 catcacctcc cacaacgagg actacaccat cgtggaacag tacgagcgcg ccgagggccg
     2641 ccactccacc ggcggcatgg acgagctgta caagaagctg aaccctcctg atgagagtgg
     2701 ccccggctgc atgagctgca agtgtgtgct ctcctgagct agcactatag tgagtcgtat
     2761 tacgtagatc cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     2821 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     2881 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     2941 ggggaggtgt gggaggtttt ttaattcgcg gccgcggcgc caatgcattg ggcccggtac
     3001 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
     3061 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     3121 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     3181 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     3241 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     3301 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     3361 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3421 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3481 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3541 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3601 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3661 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3721 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3781 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3841 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     3901 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3961 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4021 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4081 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4141 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4201 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4261 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4321 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4381 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4441 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4501 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4561 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4621 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4681 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4741 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4801 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4861 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4921 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4981 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5041 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5101 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5161 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa
     5221 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     5281 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     5341 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     5401 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     5461 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     5521 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     5581 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     5641 tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg
     5701 atcggtgcgg gcctcttcgc tattacgcca g
//
