LOCUS Exported 8213 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION mammalian bicistronic expression of mouse Olig2 and EGFP. ACCESSION . VERSION . KEYWORDS pLP-mOlig2-ires-gfp SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8213) TITLE McKinnon Lab plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 8213) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8213 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 29..133 /gene="bla" /label=AmpR promoter promoter 135..492 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(158..178) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 343..478 /label=SV40 ori /note="SV40 origin of replication" primer_bind 405..424 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 527..1321 /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(581..600) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 1191..1210 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(1509..1528) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 1553..1600 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 1929..2517 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2418..2437 /label=pBR322ori-F /note="pBR322 origin, forward primer" enhancer 2692..2995 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2996..3199 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 3150..3170 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 3266..3299 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS 5161..5196 /codon_start=1 /product="6x(His-Asn) affinity tag" /label=6xHN /translation="HNHNHNHNHNHN" terminator 5232..5326 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(5347..6006) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind 5921..5940 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" protein_bind 6035..6068 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 6203..6789 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(6370..6387) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 6597..6616 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 6790..7509 /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 6790..7506 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(6835..6856) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(7096..7115) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 7443..7464 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 7632..7753 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(7669..7688) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 7723..7742 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(7760..2) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7897..7918) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 8109..8128 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat 61 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg 121 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt 181 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 241 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 301 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg 361 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 421 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 481 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga 541 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc 601 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc 661 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc 721 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac 781 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc 841 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac 901 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg 961 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct 1021 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt 1081 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg 1141 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac 1201 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg 1261 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg 1321 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat 1381 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc 1441 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg 1501 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata 1561 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc 1621 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt 1681 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac 1741 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta 1801 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc 1861 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa 1921 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 1981 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 2041 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc 2101 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 2161 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 2221 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 2281 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 2341 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 2401 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 2461 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 2521 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc 2581 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta 2641 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata 2701 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat 2761 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga 2821 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc 2881 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt 2941 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat 3001 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag 3061 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc 3121 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga 3181 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggactca 3241 gatctcgagc tcaagcttcg aattcataac ttcgtatagc atacattata cgaagttatc 3301 agtcgacggt accggacata tgcccgggaa ttcggcacga ggccagcgga accccgaaag 3361 gtgtggatgc ttattacaga ccgagccaac accagcgccc agagccaggt tctcctccgc 3421 agcgagcacc tcaaatctaa ttcacattcg gaaggttgaa aaaagaagga tcattcgaga 3481 gcttagatca tccctggggc catggactcg gacgccagcc tggtgtctag tcgcccatcg 3541 tccccagaac ccgatgatct ttttctgccc gcccggagca aggggggaag cagcagcggc 3601 ttcacaggag ggactgtgtc ctcgtccacg ccgagcgact gcccgccaga gctgagctcc 3661 gagctacgag gggccatggg cgcgtcaggc gcgcatcccg gggacaaact gggcggcggt 3721 ggcttcaagt catcttcctc cagcacctcc tcgtccacgt cctcggcggc cacgtcttcc 3781 accaagaaag acaagaagca gatgactgag cccgagctgc agcagctgcg cctgaagatc 3841 aacagccgcg aacgcaagcg catgcacgac ctcaacatcg ccatggacgg cctgcgggag 3901 gtcatgcctt acgcgcacgg gccgtcggtg cgcaagctct ccaagatcgc cacgctgctg 3961 ctggcgcgaa actacatcct catgctcacc aactcgctgg aggagatgaa gcgactggtg 4021 agcgagatct acgggggtca ccacgccggc ttccacccat cggcctgtgg cgggctggca 4081 cactcggcgc ctctacccac cgccacggcg caccccgcgg ccgccgcgca cgccgcgcac 4141 cacccggccg tgcaccaccc catcctccct cccgcagccg ccgccgccgc cgcagccgcc 4201 gccgccgcag cggtgtccag cgcctctcta ccgggctccg ggctgtcctc ggtcggatcc 4261 atccggcctc cgcacggcct gctcaagtca ccgtcggcag ccgcagccgc ccctctcgga 4321 ggcgggggtg gaggcagcgg tggcagcggt ggcttccagc actggggcgg catgccgtgc 4381 ccctgcagca tgtgccaggt gccgccgccg caccaccacg tgtcggctat gggcgccggc 4441 accttgccgc gcctcacctc cgacgccaag tgagccggcc agcgggggtg cgtcctggag 4501 agccaagggc tgcggagcag cgcggacagg ccagcgctgg ggctccgggg ctctgttgca 4561 agaaggggag ctggaccacg ggctgggcct aggggattat ggggagtcga atatatggga 4621 accgaagcaa tgggagcatt tgaagcgatg gagagatgcg ttcgttccag aacctggttg 4681 actctttggc tgaacactcc aagggtctgg gaggcgccat tgtacaaaac ggccacaacg 4741 cgcgggtact ttcatccctg gaacttcctg cctgcacccc caggaagtct aaaaactaac 4801 tttccggtgt acaaagattg actccctgtc tgggcttaga cgagacgcag agcactgcac 4861 ttgacttctt tcccccactt caggcggggt aggagactcc caggaaccgc actgcagaag 4921 cgacctctgg gcctcaccct gcaggttcat ccctggccca gggccaggca ctaagttagt 4981 agacgctggc gtgggtatca gaagcactta gggtcctgtg gtctcagaag tgtggccttc 5041 cctcctgttg tctctcctgt tgatttgtgg tttcctaaca aatctgtgaa tgttcctttt 5101 cagagcacag gagcaagctt tctagaccat tcgtttggcg cgcgggccca ggtaagtggt 5161 cataatcata atcataatca taatcataat cacaactagc ctaggagatc ctggtcatga 5221 ctagtgcttg gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa 5281 tccagatgga gttctgaggt cattactgga tctatcaaca ggagtccaag cgagctcgat 5341 atcaaattac gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct 5401 gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac 5461 cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat 5521 attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa 5581 catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc 5641 ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga 5701 aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac 5761 cagctcaccg tctttcattg ccatacgaaa ttccggatga gcattcatca ggcgggcaag 5821 aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc 5881 cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc 5941 aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt 6001 ctccatttta gcttccttag ctcctgaaag atccataact tcgtatagca tacattatac 6061 gaagttatag atccaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 6121 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 6181 agtgccacct gacgtggatc cgcccctctc cctccccccc ccctaacgtt actggccgaa 6241 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt 6301 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg 6361 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc 6421 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc 6481 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa 6541 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc 6601 tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg 6661 gatctgatct ggggcctcgg tacacatgct ttacatgtgt ttagtcgagg ttaaaaaaac 6721 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg 6781 gccacaacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc 6841 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat 6901 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc 6961 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac 7021 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc 7081 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc 7141 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc 7201 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag 7261 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg 7321 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc 7381 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat 7441 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg 7501 tacaagtaaa gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt 7561 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa 7621 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 7681 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 7741 tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat 7801 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa 7861 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta 7921 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca 7981 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat 8041 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg 8101 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc 8161 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgc //