LOCUS       sequence-426922-        9099 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Episomal plasmid - for expression of 2 proteins linked with 
            c-terminal split intein and split puromycin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9099)
  AUTHORS   Fallahee I, Hawiger D
  TITLE     Episomal Vectors for Stable Production of Recombinant Proteins and 
            Engineered Antibodies.
  JOURNAL   Antibodies (Basel). 2024 Mar 11;13(1):18. doi: 
            10.3390/antib13010018.
  PUBMED    38534208
REFERENCE   2  (bases 1 to 9099)
  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: "Antibodies 
            (Basel)."; date: "2024-03-11"; volume: "13(1):18. doi"; pages: " 
            10.3390/antib13010018"
COMMENT     Sequence Label: 
            pCMV-protein1-IRES-protein2-IRESatt-cTermIntein-cTermPuroR
FEATURES             Location/Qualifiers
     source          1..9099
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    2228..2349
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2239..2258)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2293..2312
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2464..2510
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2540..2676
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2721..2792
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3073..3661)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3153..3172)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(5238..5306,5307..6098)
                     /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     complement(5456..5475)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6111..6247
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6424..6495
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6500..6546)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6564..6612
                     /note="synthetic polyadenylation signal"
     enhancer        6693..6996
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6998..7200
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7151..7171
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    join(7338..7911,7912..7914,7915..7923)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    join(7991..8564,8565..8567,8568..8576)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     8970..8990
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 aaaaaaaaat tgtatcaact ttctacaatc tctttcagaa gacagaagca gagggaatac
       61 ttcctaaatc attcaactag gccagcatta ccttaatacc ggaactagaa aatgacatta
      121 caagaaaaga aaacaacaga ccaatatctc tcatgaacaa agatacaaac attttcaaca
      181 aaatattagc aaaaagaatc caagaatgta tcaaaaaata tacaccacaa ccaagtagaa
      241 tttattccag atatgtaagg gtggttcaac gtttgaaaat caattaacgt aatttgtccc
      301 atcaacaggt taaagaagaa aatcacatgg tcatattgat agacacagaa aaagcatttg
      361 acaaaattta acacccattc atgatgcaat ctctcagtaa actaggaata gaggaaaact
      421 tcctcagctt gaatgtacct tcctctcaat tttgctatga acctgaaact cctcttaaaa
      481 aataaagttt ttcatttaaa aagaaaacaa aaaacatgga ggagcgttga tgtatctcat
      541 tttagaccaa tcagctatgg atagttaggc gacagcacag atagctgttg tacttctgtt
      601 tctggcaatg ttccagacta catttaaaaa atttttaatt atagacttgt acttaatgtt
      661 caagaaaaat atgaaaatgg ctttgccgtg ttaatgctac tcttttttaa aaaaaactaa
      721 agttcaaact ttatttatat ttcattagtt ttttagctac tgttcttttt ctgttctggg
      781 atctcattca gaatgccaca ttacatataa ttctcatgtc tccttgggtt cctcttagtt
      841 ttgacagttc ctcagacttt tcttattttt gatgaccttg acagttttga ggagtactgg
      901 ttagatatag ggtaatggtt tttaaagtat atttgtcatg atttatactg gggtaagggt
      961 ttggggagga agcccatggg gtaaagtact gttctcatca catcatatca aggttatata
     1021 ccatcaatat tgccacagat gttacttagc cttttaatat ttctctaatt tagtgtatat
     1081 gcaatgatag ttctctgatt tctgagattg agtttctcat gtgtaatgat tatttagagt
     1141 ttctctttca tctgttcaaa tttttgtcta gttttatttt ttactgattt gtaagacttc
     1201 tttttataat ctgcatatta caattctctt tactggggtg ttgcaaatat tttctgtcat
     1261 tctatggcct gacttttctt aatggttttt taattttaaa aataagtctt aatattcatg
     1321 caatctaatt aacaatcttt tctttgtggt taggactttg agtcataaga aatttttctc
     1381 tacactgaag tcatgatggc atgcttctat attattttct aaaagattta aagttttgcc
     1441 ttctccattt agacttataa ttcactggaa tttttttgtg tgtatggtat gacatatggg
     1501 ttccctttta ttttttacat ataaatatat ttccctgttt ttctaaaaaa gaaaaagatc
     1561 atcattttcc cattgtaaaa tgccatattt ttttcatagg tcacttacat atatcaatgg
     1621 gtctgtttct gagctctact ctattttatc agcctcactg tctatcccca cacatctcat
     1681 gctttgctct aaatcttgat atttagtgga acattctttc ccattttgtt ctacaagaat
     1741 atttttgtta ttgtcttttg ggcttctata tacattttag aatgaggttg gcaagttaac
     1801 aaacagcttt tttggggtga acatattgac tacaaattta tgtggaaaga aagtatacct
     1861 tcacaatatt aagtctttta gttcatgaat atagtatgtc tctccgtttc tgcattaact
     1921 tagacattca ttaatttctc tcacaattta taagtttatt tagatcttca ttcatttaaa
     1981 tcttcactaa cctctcattt acaatttgta agttttctgg gtaacagtct tgcacttctt
     2041 tgcctagatt tatttccaag tagattattt tcatacatcg tctatggtgt catttttaaa
     2101 atgtaatttt tcaccttttt attgctaaag agagatgact gattgttaat attgatcttg
     2161 tgcgtggcga ccttgctgtc tagattgact gactgagata cagcgtacct tcagctcaca
     2221 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     2281 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     2341 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     2401 gaggtttttt aaagcaagta aaacctctac aaatgtggta ttggcccatc tctatcggta
     2461 tcgtagcata accccttggg gcctctaaac gggtcttgag gggttttttg tgcccctcgg
     2521 gccggattgc tatctaccgg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt
     2581 cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt
     2641 ccatacgtgt cctccttacc agaaatttat ccttaaggtc gtcagctatc ctgcgagcga
     2701 tctctcgatt tcgatcaaga cattccttta atggtctttt ctggacacca ctaggggtca
     2761 gaagtagttc atcaaacttt cttccctccc taatctcaat cgcgaactaa tttgcgctgc
     2821 tggcgcatga agaataggac ttgtctgggt acgagcgtca gcctgcaggt gcctaactta
     2881 attaaccagt caagtcagct acttggcgag atcgacttgt ctgggtttcg actacgctca
     2941 gaattgcgtc agtcaagttc gatctggtcc ttgctattgc acccgttctc cgattacgag
     3001 tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3061 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3121 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3181 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3241 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3301 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3361 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3421 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3481 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3541 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3601 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3661 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3721 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3781 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3841 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcatt
     3901 taaatttccg aactctccaa ggccctcgtc ggaaaatctt caaacctttc gtccgatcca
     3961 tcttgcaggc tacctctcga acgaactatc gcaagtctct tggccggccg cattttcttt
     4021 gacccaggat atttgcaaaa gacatattca aacttccgca gaacacttta tttcacatat
     4081 acatgcctct tatatcaggg atgtgaaaca gggtcttgaa aactgtctaa atctaaaaca
     4141 atgctaatgc aggtttaaat ttaataaaat aaaatccaaa atctaacagc caagtcaaat
     4201 ctgtatgttt taacatttaa aatattttaa agacgtcttt tcccaggatt caacatgtga
     4261 aatcttttct cagggataca cgtgtgccta gatcctcatt gctttagttt tttacagagg
     4321 aatgaatata aaaagaaaat acttaaattt tatccctctt acctctataa tcatacatag
     4381 gcataatttt ttaacctagg ctccagatag ccatagaaga accaaacact ttctgcgtgt
     4441 gtgagaataa tcagagtgag attttttcac aagtacctga tgagggttga gacaggtaga
     4501 aaaagtgaga gatctctatt tatttagcaa taatagagaa agcatttaag agaataaagc
     4561 aatggaaata agaaatttgt aaatttcctt ctgataacta gaaatagagg atccagtttc
     4621 ttttggttaa cctaaatttt atttcatttt attgttttat tttattttat tttattttat
     4681 tttgtgtaat cgtagtttca gagtgttaga gctgaaagga agaagtagga gaaacatgca
     4741 aagtaaaagt ataacacttt ccttactaaa ccgacatggg tttccaggta ggggcaggat
     4801 tcaggatgac tgacagggcc cttagggaac actgagaccc tacgctgacc tcataaatgc
     4861 ttgctacctt tgctgtttta attacatctt ttaatagcag gaagcagaac tctgcacttc
     4921 aaaagttttt cctcacctga ggagttaatt tagtacaagg ggaaaaagta cagggggatg
     4981 ggggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg
     5041 ttgcttggca gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat
     5101 attgtactca agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt
     5161 ctgatcggtc aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat
     5221 cagcaggagg ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5281 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5341 aagatcagtt gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5401 ttgagagttt tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat
     5461 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5521 attctcagaa tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca
     5581 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5641 tacttctgac aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5701 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5761 agcgtgacac cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg
     5821 aactacttac tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg
     5881 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5941 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     6001 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     6061 tcgctgagat aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc
     6121 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     6181 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     6241 tccttaagat cccgaatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg
     6301 agcttacgcg aacagccgtg gcgctcattt gctcgtcatt gcgaactaat ttgcgctgct
     6361 ggcgcatgaa gaatagtatc acgcgtgtca gcttaccttt ttggcagcga tcgcggctcc
     6421 cgacatcttg gaccattagc tccacaggta tcttcttccc tctagtggtc ataacagcag
     6481 cttcagctac ctctcaattc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt
     6541 atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca cacatccatc ttcgatggat
     6601 agcgatttta ttatctaact gctgatcgag tgtagccaga tctagtaatc aattacgggg
     6661 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     6721 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     6781 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     6841 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     6901 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     6961 cagtacatct acgtattagt catcgctatt accatgctga tgcggttttg gcagtacatc
     7021 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     7081 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     7141 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     7201 ggtttagtga accgtcagat cagatctttg tcgatcctac catccactcg acacacccgc
     7261 cagcggccgc gccgccacca tgtagattgc gaactaagaa ttctaaggat ccaatcttct
     7321 actgtcgtat gccataaccc ctctccctcc ccccccccta acgttactgg ccgaagccgc
     7381 ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt
     7441 ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt
     7501 tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg
     7561 gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca
     7621 cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg
     7681 gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc
     7741 tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct
     7801 gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaacgtctag
     7861 gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca
     7921 accatggatt agtttgcgct gctggcgcat gaagaataag ctagcttaat gccataaagc
     7981 gaatacgatt cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat
     8041 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     8101 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     8161 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     8221 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     8281 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     8341 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     8401 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     8461 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaacgtc taggcccccc
     8521 gaaccacggg gacgtggttt tcctttgaaa aacacgatga taagcttgcc acaaccccgg
     8581 gagatgagga tcgtttcgca tgatgctgaa aaaaatcctg aaaatcgaag agctggatga
     8641 acgtgaactg atcgatattg aggtgtccgg taaccacctg ttttacgcta acgatattct
     8701 gacccacaac tcttcttctg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     8761 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     8821 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     8881 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     8941 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     9001 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgg acctggtgca tgacccgcaa
     9061 gcccggtgcc tgataagtcg acctatcaaa tatttaaag
//
