LOCUS Exported 7579 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION For transfection or retroviral production. Mammalian expression of EGFP driven by the ECMV IRES.. ACCESSION . VERSION . KEYWORDS pLPCX-IRES-GFP 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7579 /organism="synthetic DNA construct" /mol_type="other DNA" 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 2946..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 3533..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 3533..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 complement(5164..5197) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind complement(5257..5290) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." 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(6340..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 //