LOCUS       sequence-141470-        5728 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCS4+-NLS-EGFP-T2A-mCherry-CAAX.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5728)
  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 5728)
  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-102
FEATURES             Location/Qualifiers
     source          1..5728
                     /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..1951
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1961..2668
                     /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(2106..2124)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2374..2393
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2568..2591
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2740..2759)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2741..2759)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2764..2898
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2788..2807)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2842..2861
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3011..3031)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3011..3029)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3050..3066)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3050..3066)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3063..3085)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3074..3090
                     /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(3098..3104,3105..3122,3123..3128))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3143..3164
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3281..3298)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3452..4040)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3532..3551)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4211..5002,5003..5071))
                     /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     4834..4853
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5072..5176)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5202..5657)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5339..5360)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5551..5570
                     /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 aagcggagag ggcagaggaa gtctgctaac
     1921 atgcggtgac gtcgaggaga atcctggacc tagatctacc atggtgagca agggcgagga
     1981 ggacaacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt
     2041 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
     2101 gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc
     2161 ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta
     2221 cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     2281 cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt
     2341 gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg
     2401 ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa
     2461 gcagaggctg aagctgaagg acggcggcca ctacgacgcc gaggtcaaga ccacctacaa
     2521 ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagc tggacatcac
     2581 ctcccacaac gaggactaca ccatcgtgga acagtacgag cgcgccgagg gccgccactc
     2641 caccggcggc atggacgagc tgtacaagaa gctgaaccct cctgatgaga gtggccccgg
     2701 ctgcatgagc tgcaagtgtg tgctctcctg agctagcctc cctatagtga gtcgtattac
     2761 gtagatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     2821 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     2881 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     2941 gaggtgtggg aggtttttta attcgcggcc gcggcgccaa tgcattgggc ccggtaccca
     3001 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
     3061 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     3121 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     3181 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     3241 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     3301 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3361 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3421 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3481 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3541 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3601 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3661 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3721 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3781 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3841 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     3901 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3961 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4021 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4081 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4141 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4201 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4261 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     4321 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     4381 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     4441 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4501 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4561 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4621 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4681 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4741 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4801 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4861 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4921 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4981 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5041 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     5101 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     5161 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     5221 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     5281 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     5341 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     5401 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     5461 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     5521 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     5581 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     5641 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc
     5701 ggtgcgggcc tcttcgctat tacgccag
//
