LOCUS       sequence-141474-        5000 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCS4+-P2A-mCherry-CAAX.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5000)
  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 5000)
  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-187
FEATURES             Location/Qualifiers
     source          1..5000
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             180..236
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             246..953
                     /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(391..409)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     659..678
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     853..876
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1025..1044)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1026..1044)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1049..1183
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1073..1092)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1127..1146
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1296..1316)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1296..1314)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1335..1351)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1335..1351)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1348..1370)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1359..1375
                     /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(1383..1389,1390..1407,1408..1413))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1428..1449
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1566..1583)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1737..2325)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1817..1836)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2496..3287,3288..3356))
                     /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     3119..3138
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3357..3461)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3487..3942)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3624..3645)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3836..3855
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4016..5000
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttgtacagat atctggcgcg cctaccatgg
      121 ggcccggccg gcctaagatc accggtcccg ggcttaagga gctcgcatgc ggaagcggag
      181 ctactaactt cagcctgctg aagcaggctg gagacgtgga ggagaaccct ggacctagat
      241 ctaccatggt gagcaagggc gaggaggaca acatggccat catcaaggag ttcatgcgct
      301 tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg
      361 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggcggccccc
      421 tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga
      481 agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg
      541 agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc
      601 aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc
      661 ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg
      721 acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg
      781 acgccgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca
      841 acgtcaacat caagctggac atcacctccc acaacgagga ctacaccatc gtggaacagt
      901 acgagcgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac aagaagctga
      961 accctcctga tgagagtggc cccggctgca tgagctgcaa gtgtgtgctc tcctgagcta
     1021 gcctccctat agtgagtcgt attacgtaga tccagacatg ataagataca ttgatgagtt
     1081 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     1141 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     1201 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaattcg cggccgcggc
     1261 gccaatgcat tgggcccggt acccagcttt tgttcccttt agtgagggtt aattgcgcgc
     1321 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     1381 cacaacatac gagccgggag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     1441 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     1501 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     1561 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     1621 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     1681 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     1741 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     1801 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     1861 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     1921 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     1981 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2041 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     2101 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     2161 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     2221 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     2281 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     2341 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     2401 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     2461 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     2521 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     2581 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     2641 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     2701 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     2761 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     2821 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     2881 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     2941 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     3001 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     3061 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     3121 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     3181 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     3241 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     3301 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     3361 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     3421 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     3481 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     3541 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     3601 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     3661 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     3721 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     3781 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     3841 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     3901 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
     3961 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagtcgacca
     4021 tagccaattc aatatggcgt atatggactc atgccaattc aatatggtgg atctggacct
     4081 gtgccaattc aatatggcgt atatggactc gtgccaattc aatatggtgg atctggaccc
     4141 cagccaattc aatatggcgg acttggcacc atgccaattc aatatggcgg acttggcact
     4201 gtgccaactg gggaggggtc tacttggcac ggtgccaagt ttgaggaggg gtcttggccc
     4261 tgtgccaagt ccgccatatt gaattggcat ggtgccaata atggcggcca tattggctat
     4321 atgccaggat caatatatag gcaatatcca atatggccct atgccaatat ggctattggc
     4381 caggttcaat actatgtatt ggccctatgc catatagtat tccatatatg ggttttccta
     4441 ttgacgtaga tagcccctcc caatgggcgg tcccatatac catatatggg gcttcctaat
     4501 accgcccata gccactcccc cattgacgtc aatggtctct atatatggtc tttcctattg
     4561 acgtcatatg ggcggtccta ttgacgtata tggcgcctcc cccattgacg tcaattacgg
     4621 taaatggccc gcctggctca atgcccattg acgtcaatag gaccacccac cattgacgtc
     4681 aatgggatgg ctcattgccc attcatatcc gttctcacgc cccctattga cgtcaatgac
     4741 ggtaaatggc ccacttggca gtacatcaat atctattaat agtaacttgg caagtacatt
     4801 actattggaa gtacgccagg gtacattggc agtactccca ttgacgtcaa tggcggtaaa
     4861 tggcccgcga tggctgccaa gtacatcccc attgacgtca atggggaggg gcaatgacgc
     4921 aaatgggcgt tccattgacg taaatgggcg gtaggcgtgc ctaatgggag gtctatataa
     4981 gcaatgctcg tttagggaac
//
