LOCUS       sequence-110889-        7579 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For transfection or retroviral production. Mammalian expression of 
            EGFP driven by the ECMV IRES..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7579)
  AUTHORS   Salat-Canela C, Sese M, Peula C, Ramon y Cajal S, Aasen T
  TITLE     Internal translation of the connexin 43 transcript.
  JOURNAL   Cell Commun Signal. 2014 May 8;12:31. doi: 10.1186/1478-811X-12-31.
  PUBMED    24884945
REFERENCE   2  (bases 1 to 7579)
  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: "Cell Commun
            Signal."; date: "2014-05-8"; volume: "12:31. doi"; pages: " 
            10.1186/1478-811X-12-31"
COMMENT     Sequence Label: pLPCX-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..7579
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             414..589
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    652..851
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1052..1468
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1398..1420
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1436..1452
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1566..2165
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     primer_bind     complement(1566..1585)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2062..2082
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     enhancer        2340..2643
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2644..2847
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2798..2818
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    2888..2944
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    join(2946..3520,3521..3523,3524..3532)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3113..3130)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3340..3359
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3533..3535,3536..3538,3539..4252)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3533..3535,3536..3538,3539..4249)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3578..3599)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3839..3858)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4186..4207
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4318..4911
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5014..5036)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     protein_bind    5164..5197
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    5257..5290
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     5310..5329
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5410..5427)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5581..6169)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5661..5680)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6340..7131,7132..7200))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6963..6982
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7201..7305)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7373..7391
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tttgaaagac cccacccgta ggtggcaagc tagcttaagt aacgccactt tgcaaggcat
       61 ggaaaaatac ataactgaga atagaaaagt tcagatcaag gtcaggaaca aagaaacagc
      121 tgaataccaa acaggatatc tgtggtaagc ggttcctgcc ccggctcagg gccaagaaca
      181 gatgagacag ctgagtgatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
      241 ctcggggcca agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagtgaa
      301 tcatcagatg tttccagggt gccccaagga cctgaaaatg accctgtacc ttatttgaac
      361 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc cgctctccga gctcaataaa
      421 agagcccaca acccctcact cggcgcgcca gtcttccgat agactgcgtc gcccgggtac
      481 ccgtattccc aataaagcct cttgctgttt gcatccgaat cgtggtctcg ctgttccttg
      541 ggagggtctc ctctgagtga ttgactaccc acgacggggg tctttcattt gggggctcgt
      601 ccgggatttg gagacccctg cccagggacc accgacccac caccgggagg taagctggcc
      661 agcaacttat ctgtgtctgt ccgattgtct agtgtctatg tttgatgtta tgcgcctgcg
      721 tctgtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt
      781 ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg
      841 cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt
      901 aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa
      961 ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct
     1021 gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt
     1081 gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa
     1141 gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc
     1201 gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc
     1261 tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt
     1321 tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc
     1381 atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta
     1441 tccagccctc actccttctc taggcgccgg aattgatccc ccgggctgca ggtcggccgc
     1501 cacgaccggt gccgccacca tcccctgacc cacgcccctg acccctcaca aggagacgac
     1561 cttccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg
     1621 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc
     1681 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     1741 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     1801 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     1861 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     1921 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     1981 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     2041 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     2101 ccgccgacgt cgagtgcccg aaggaccgcg cgacctggtg catgacccgc aagcccggtg
     2161 cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga ccccatggct
     2221 ccgaccgaag ccgacccggg cggccccgcc gaccccgcac ccgcccccga ggcccaccga
     2281 ctctagtaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     2341 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     2401 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     2461 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     2521 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     2581 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     2641 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     2701 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     2761 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     2821 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac
     2881 cggactcaga tctcgagctc aagcttcgaa ttctgcagtc gacggtaccg cgggcccggg
     2941 atccgcccct ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     3001 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     3061 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     3121 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
     3181 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     3241 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     3301 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     3361 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     3421 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac
     3481 cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatggtgag
     3541 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     3601 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     3661 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     3721 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     3781 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     3841 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     3901 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     3961 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     4021 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     4081 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     4141 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     4201 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccg
     4261 cctcggccaa acatcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     4321 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     4381 gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagc
     4441 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     4501 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     4561 cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga
     4621 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
     4681 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
     4741 aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt
     4801 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt
     4861 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcat ttgggggctc
     4921 gtccgggatc gggagacccc tgcccaggga ccaccgaccc accaccggga ggtaagctgg
     4981 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     5041 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     5101 gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta
     5161 ctgataactt cgtataatgt atgctatacg aagttattag gtctgaagag gagtttacgt
     5221 ccagccaagc tagggccgcg atccggaacc cttaatataa cttcgtataa tgtatgctat
     5281 acgaagttat cagtactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     5341 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc
     5401 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5461 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5521 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5581 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5641 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5701 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5761 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5821 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5881 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5941 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6001 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     6061 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6121 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6181 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6241 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6301 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6361 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6421 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6481 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6541 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6601 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctgcagg
     6661 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6721 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6781 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6841 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6901 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg
     6961 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     7021 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7081 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7141 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7201 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7261 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7321 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     7381 tatcacgagg ccctttcgtc ttcaagaatt gctagcaatt gctagcaatt gctagcaatt
     7441 cataccagat caccgaaaac tgtcctccaa atgtgtcccc ctcacactcc caaattcgcg
     7501 ggcttctgcc tcttagacca ctctacccta ttccccacac tcaccggagc caaagccgcg
     7561 gcccttccgt ttctttgct
//
