LOCUS       sequence-12670-e        7110 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7110)
  AUTHORS   Trichas G, Begbie J, Srinivas S
  TITLE     Use of the viral 2A peptide for bicistronic expression in transgenic
            mice.
  JOURNAL   BMC Biol. 2008 Sep 15;6:40. doi: 10.1186/1741-7007-6-40.
  PUBMED    18793381
REFERENCE   2  (bases 1 to 7110)
  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: "BMC Biol.";
            date: "2008-09-15"; volume: "6:40. doi"; pages: " 
            10.1186/1741-7007-6-40"
COMMENT     Sequence Label: pCS2-TAG
FEATURES             Location/Qualifiers
     source          1..7110
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..67)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(47..65)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(86..102)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(86..102)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(99..121)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    110..126
                     /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(134..140,141..158,159..164))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    179..200
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(317..334)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(488..1076)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(568..587)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1247..2038,2039..2107))
                     /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     1870..1889
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2108..2212)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2238..2693)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2375..2396)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2587..2606
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2767..3751
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        3786..3804
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3786..3803
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      3872..3881
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3926..3935
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3995..5422
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     CDS             5432..5485
                     /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             5504..5881
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAVTKYTSAK"
     CDS             join(5966..5968,5969..6679)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(6008..6029)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6269..6288)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6616..6637
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    6964..7098
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7001..7020)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 gcggccgcgg cgccaatgca ttgggcccgg tacccagctt ttgttccctt tagtgagggt
       61 taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
      121 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
      181 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
      241 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
      301 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
      361 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
      421 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
      481 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
      541 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
      601 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
      661 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
      721 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
      781 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
      841 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
      901 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
      961 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     1021 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     1081 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     1141 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     1201 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     1261 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     1321 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     1381 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     1441 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     1501 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     1561 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     1621 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     1681 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     1741 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     1801 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     1861 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     1921 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     1981 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     2041 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     2101 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     2161 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     2221 cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     2281 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     2341 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     2401 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     2461 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     2521 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     2581 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     2641 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc
     2701 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     2761 ccagtcgacc atagccaatt caatatggcg tatatggact catgccaatt caatatggtg
     2821 gatctggacc tgtgccaatt caatatggcg tatatggact cgtgccaatt caatatggtg
     2881 gatctggacc ccagccaatt caatatggcg gacttggcac catgccaatt caatatggcg
     2941 gacttggcac tgtgccaact ggggaggggt ctacttggca cggtgccaag tttgaggagg
     3001 ggtcttggcc ctgtgccaag tccgccatat tgaattggca tggtgccaat aatggcggcc
     3061 atattggcta tatgccagga tcaatatata ggcaatatcc aatatggccc tatgccaata
     3121 tggctattgg ccaggttcaa tactatgtat tggccctatg ccatatagta ttccatatat
     3181 gggttttcct attgacgtag atagcccctc ccaatgggcg gtcccatata ccatatatgg
     3241 ggcttcctaa taccgcccat agccactccc ccattgacgt caatggtctc tatatatggt
     3301 ctttcctatt gacgtcatat gggcggtcct attgacgtat atggcgcctc ccccattgac
     3361 gtcaattacg gtaaatggcc cgcctggctc aatgcccatt gacgtcaata ggaccaccca
     3421 ccattgacgt caatgggatg gctcattgcc cattcatatc cgttctcacg ccccctattg
     3481 acgtcaatga cggtaaatgg cccacttggc agtacatcaa tatctattaa tagtaacttg
     3541 gcaagtacat tactattgga aggacgccag ggtacattgg cagtactccc attgacgtca
     3601 atggcggtaa atggcccgcg atggctgcca agtacatccc cattgacgtc aatggggagg
     3661 ggcaatgacg caaatgggcg ttccattgac gtaaatgggc ggtaggcgtg cctaatggga
     3721 ggtctatata agcaatgctc gtttagggaa ccgccattct gcctggggac gtcggagcaa
     3781 gcttgattta ggtgacacta tagaatacaa gctacttgtt ctttttgcag gatcccatcg
     3841 attcgaattc aaggcctctc gagcctctag agccaccatg ggctgcatca agagcaagcg
     3901 caaggacaac ctgaacgacg acgaggccgc catgggctgc atcaagagca agcgcaagga
     3961 caacctgaac gacgacgagg caccggtcgc ctccatggtg agcaagggcg aggaggtcat
     4021 caaagagttc atgcgcttca aggtgcgcat ggagggctcc atgaacggcc acgagttcga
     4081 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     4141 gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tccccccagt tcatgtacgg
     4201 ctccaaggcg tacgtgaagc accccgccga catccccgat tacaagaagc tgtccttccc
     4261 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggtctgg tgaccgtgac
     4321 ccaggactcc tccctgcagg acggcacgct gatctacaag gtgaagatgc gcggcaccaa
     4381 cttccccccc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccaccga
     4441 gcgcctgtac ccccgcgacg gcgtgctgaa gggcgagatc caccaggccc tgaagctgaa
     4501 ggacggcggc cactacctgg tggagttcaa gaccatctac atggccaaga agcccgtgca
     4561 actgcccggc tactactacg tggacaccaa gctggacatc acctcccaca acgaggacta
     4621 caccatcgtg gaacagtacg agcgctccga gggccgccac cacctgttcc tggggcatgg
     4681 caccggcagc accggcagcg gcagctccgg caccgcctcc tccgaggaca acaacatggc
     4741 cgtcatcaaa gagttcatgc gcttcaaggt gcgcatggag ggctccatga acggccacga
     4801 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     4861 gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc cccagttcat
     4921 gtacggctcc aaggcgtacg tgaagcaccc cgccgacatc cccgattaca agaagctgtc
     4981 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gtctggtgac
     5041 cgtgacccag gactcctccc tgcaggacgg cacgctgatc tacaaggtga agatgcgcgg
     5101 caccaacttc ccccccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     5161 caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccacc aggccctgaa
     5221 gctgaaggac ggcggccact acctggtgga gttcaagacc atctacatgg ccaagaagcc
     5281 cgtgcaactg cccggctact actacgtgga caccaagctg gacatcacct cccacaacga
     5341 ggactacacc atcgtggaac agtacgagcg ctccgagggc cgccaccacc tgttcctgta
     5401 cggcatggac gagctgtaca agggcagtgg agagggcaga ggaagtctgc taacatgcgg
     5461 tgacgtcgag gagaatcctg gcccaggaga tctgaatatc accatgccag agccagcgaa
     5521 gtctgctccc gccccgaaaa agggctccaa gaaggcggtg actaaggcgc agaagaaagg
     5581 cggcaagaag cgcaagcgca gccgcaagga gagctattcc atctatgtgt acaaggttct
     5641 gaagcaggtc caccctgaca ccggcatttc gtccaaggcc atgggcatca tgaattcgtt
     5701 tgtgaacgac attttcgagc gcatcgcagg tgaggcttcc cgcctggcgc attacaacaa
     5761 gcgctcgacc atcacctcca gggagatcca gacggccgtg cgcctgctgc tgcctgggga
     5821 gttggccaag cacgccgtgt ccgagggtac taaggccgtc accaagtaca ccagcgctaa
     5881 gggatctggg ggtggaggct ccggcggggg tggatctgga ggtgggggca gcggcggagg
     5941 tgggctagcg ctaccgcggg ccaccatgag caagggcgag gagctgttca ccggggtggt
     6001 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     6061 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     6121 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     6181 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     6241 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     6301 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     6361 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     6421 catggccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga
     6481 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     6541 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     6601 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactcacgg
     6661 catggacgag ctgtacaagt ccggactcag atcatcaagc ttatcggatc cgggcggccg
     6721 tgataatagg tgatagtaag taatagtgag atatctctag aattccagct gagcgccggt
     6781 cgctaccatt accagttggt ctggtgtcaa aaataataat aaccgggcag gggggatcta
     6841 agctctagat aagtaatgat cataatcagc catatcacat ctgtagaggt tttacttgct
     6901 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     6961 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     7021 acaaataaag catttttttc actgcattct agttgtggtt tgtccaanct catcaatgta
     7081 tcttatcatg tctggatcca ctagttcaga
//
