LOCUS Exported 5795 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Retroviral vector with N terminal EGFP. ACCESSION . VERSION . KEYWORDS pBABE puro EGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5795) TITLE Emory Custom Cloning Core Plasmids - Oskar Laur REFERENCE 2 (bases 1 to 5795) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5795 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 8..477 /note="long terminal repeat from Moloney murine leukemia virus" misc_feature 542..741 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 942..1358 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1288..1310 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1326..1342 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" regulatory 1369..1378 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1375..2091 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 1375..2091 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1420..1441) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1681..1700) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2028..2049 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(2125..2145) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2130..2459 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2310..2445 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2372..2391 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2469..3068 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(2469..2488) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 2965..2985 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 3289..3758 /note="long terminal repeat from Moloney murine leukemia virus" primer_bind complement(3888..3905) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4059..4647) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4139..4158) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4807..5667) /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 5430..5449 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5668..5772) /gene="bla" /label=AmpR promoter ORIGIN 1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca 61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt 121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag 181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc 241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga 301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc 361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc 421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt 481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg 541 caagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta 601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa 661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc 721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg 781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt 841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt 901 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc 961 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg 1021 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg 1081 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg 1141 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag 1201 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc 1261 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat 1321 cctcccttta tccagccctc actccttctc taggcgccgg ccggatccgc caccatggtg 1381 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac 1441 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag 1501 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg 1561 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac 1621 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag 1681 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac 1741 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg 1801 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc 1861 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac 1921 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg 1981 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg 2041 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa ggtaggaatt 2101 cgccagcaca gtggtcgacc ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag 2161 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg 2221 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 2281 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc 2341 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg 2401 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa 2461 agcttaccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca 2521 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg 2581 atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg 2641 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca 2701 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt 2761 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc 2821 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg 2881 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg 2941 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg 3001 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg 3061 gtgcctgacg cccgccccac gacccgcagc gcccgaccga aaggagcgca cgaccccatg 3121 catcgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca 3181 cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaaatacat 3241 aactgagaat agagaagttc agatcaaggt caggaacaga tggaacagct gaatatgggc 3301 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa 3361 cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc 3421 caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga 3481 tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc 3541 agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac 3601 aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc 3661 caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc 3721 ctctgagtga ttgactaccc gtcagcgggg gtctttcaca tgcagcatgt atcaaaatta 3781 atttggtttt ttttcttaag tatttacatt aaatggccat agttgcatta atgaatcggc 3841 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac 3901 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata 3961 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa 4021 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 4081 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 4141 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 4201 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca 4261 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 4321 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 4381 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 4441 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga 4501 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 4561 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 4621 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 4681 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc 4741 ttcacctaga tccttttgcg gccgcaaatc aatctaaagt atatatgagt aaacttggtc 4801 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 4861 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 4921 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 4981 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 5041 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 5101 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 5161 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 5221 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 5281 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 5341 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 5401 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 5461 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 5521 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 5581 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 5641 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 5701 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 5761 aggggttccg cgcacatttc cccgaaaagt gccac //