LOCUS       sequence-142788-        5992 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  based on pCX-C1 with F2A1/F2A2 inserted.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5992)
  AUTHORS   De Giorgi M, Cinti A, Pelikant-Malecka I, Chisci E, Lavitrano M, 
            Giovannoni R, Smolenski RT
  TITLE     Co-expression of functional human Heme Oxygenase 1, 
            Ecto-5'-Nucleotidase and ecto-nucleoside triphosphate 
            diphosphohydrolase-1 by ""self-cleaving"" 2A peptide system.
  JOURNAL   Plasmid. 2015 May;79:22-9. doi: 10.1016/j.plasmid.2015.03.004. Epub 
            2015 Mar 14.
  PUBMED    25779031
REFERENCE   2  (bases 1 to 5992)
  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: "Plasmid."; 
            date: "2015-05"; volume: "79:22-9. doi"; pages: " 
            10.1016/j.plasmid.2015.03.004. Epub 2015 Mar 14"
COMMENT     Sequence Label: pCX-2A-2A
FEATURES             Location/Qualifiers
     source          1..5992
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        15..394
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        397..672
                     /label=chicken beta-actin promoter
     intron          673..1685
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1609..1631
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1693..1712
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1746..1811
                     /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             1839..1904
                     /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"
     primer_bind     complement(2003..2022)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2068..2123
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2122..2141)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2484..2500)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2484..2500)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2497..2519)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2508..2524
                     /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(2532..2538,2539..2556,2557..2562))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2577..2598
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     polyA_signal    2780..2901
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2817..2836)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2871..2890
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2908..3363)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3045..3066)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3257..3276
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3390..3494
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3496..3853
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3496..3687
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3519..3539)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3704..3839
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3888..4682
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3888..4682
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3942..3961)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4552..4571
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4870..4889)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4914..4961
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5290..5878
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5779..5798
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattat ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
       61 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      121 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      181 aacgccaata gggactttcc attgacgtca atgggtggac tatttacggt aaactgccca
      241 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      301 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      361 gtacatctac gtattagtca tcgctattac catgggtcga ggtgagcccc acgttctgct
      421 tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt attttttaat
      481 tattttgtgc agcgatgggg gcgggggggg ggggggcgcg cgccaggcgg ggcggggcgg
      541 ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg
      601 ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc
      661 gcgcggcggg cgggagtcgc tgcgttgcct tcgccccgtg ccccgctccg cgccgcctcg
      721 cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc
      781 ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac ggctcgtttc ttttctgtgg
      841 ctgcgtgaaa gccttaaagg gctccgggag ggccctttgt gcggggggga gcggctcggg
      901 gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc ggcccgcgct gcccggcggc
      961 tgtgagcgct gcgggcgcgg cgcggggctt tgtgcgctcc gcgtgtgcgc gaggggagcg
     1021 cggccggggg cggtgccccg cggtgcgggg gggctgcgag gggaacaaag gctgcgtgcg
     1081 gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg gcggtcgggc tgtaaccccc
     1141 ccctgcaccc ccctccccga gttgctgagc acggcccggc ttcgggtgcg gggctccgta
     1201 cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg
     1261 gcggggcggg gccgcctcgg gccggggagg gctcggggga ggggcgcggc ggcccccgga
     1321 gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg taatcgtgcg
     1381 agagggcgca gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc
     1441 gccgcacccc ctctagcggg cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg
     1501 gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc cagcctcggg
     1561 gctgtccgcg gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct
     1621 ggcgtgtgac cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc
     1681 tacagctcct gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaattcg
     1741 gtaccgtgaa acagactttg aattttgacc ttctcaagtt ggcgggagac gtggagtcca
     1801 acccagggcc cggcagcggc gctagcgaat tcggtaccgt gaaacagact ttgaattttg
     1861 accttctcaa gttggcggga gacgtggagt ccaacccagg gcccggcagc ggcgctagcg
     1921 aattcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc
     1981 tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga
     2041 agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt
     2101 gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac
     2161 atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga
     2221 acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct
     2281 tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat
     2341 ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc
     2401 tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacctgc
     2461 agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     2521 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     2581 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     2641 tcgtgccagc ggatccaccg gatctagata actgatcata atcagccata ccacatttgt
     2701 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     2761 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     2821 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     2881 caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     2941 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     3001 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     3061 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     3121 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     3181 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     3241 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     3301 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     3361 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3421 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3481 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     3541 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3601 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3661 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3721 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3781 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3841 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     3901 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     3961 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4021 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     4081 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4141 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4201 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4261 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4321 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4381 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     4441 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4501 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4561 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4621 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4681 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     4741 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4801 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     4861 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     4921 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     4981 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     5041 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     5101 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     5161 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5221 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5281 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5341 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5401 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     5461 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5521 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5581 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5641 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5701 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5761 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5821 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5881 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     5941 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc at
//
