LOCUS Exported 8665 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pIRES2-eGFP-Ofd1-Myc SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8665) AUTHORS Singla V, Hunkapiller J, Santos N, Seol AD, Norman AR, Wakenight P, Skarnes WC, Reiter JF TITLE Floxin, a resource for genetically engineering mouse ESCs. JOURNAL Nat Methods. 2010 Jan . 7(1):50-2. PUBMED 19966808 REFERENCE 2 (bases 1 to 8665) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8665 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 3943..3972 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" misc_feature 4024..4610 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4191..4208) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 4418..4437 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 4611..5330 /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 4611..5327 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(4656..4677) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4917..4936) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 5264..5285 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 5453..5574 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5490..5509) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5544..5563 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(5581..6036) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5718..5739) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5930..5949 /label=F1ori-R /note="F1 origin, reverse primer" promoter 6063..6167 /gene="bla" /label=AmpR promoter promoter 6169..6526 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(6192..6212) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 6377..6512 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6439..6458 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 6561..7355 /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(6615..6634) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 7225..7244 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(7543..7562) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 7587..7634 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 7963..8551 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8452..8471 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 601 ccggactcag atctcgagct caagcttcga attatctgca gaattcgccc tttccgtctt 661 tattccgtcc agagtccctc ctggctcgcg gacccgccca cccttcccat accgtgggag 721 gaagcgcggc tgcttgacga gttaggggct gaatgtttag ctctcttgct cccctaggtc 781 cccagagaca agctggattc cgaggtcaag ggctccctct cctcctgcgc cccggcccgg 841 gccgaggagt ccgaccgtgg aggggcgagc tcgggaggaa agcagatgag gatggctcag 901 tccaacatgc cccacaagtc tgatgtgttg agtcaagatg aactgcgcaa aaagctatac 961 cagacgttta aggatcgggg tgtactggac acacttcaga cccaactccg aaaccagcta 1021 attcatgaat tgatgcatcc tgtattgagt ggagaagtga agccgccatc catttcagtg 1081 gaagggagtg ccctcctaat aggggcttct aactcgctgg tggcagacca cttacaaaga 1141 tgtggctatg aatattcact ctctgttttc tttccagaaa gtggtttggc aaaagaaaag 1201 atatttacta tgcaggatct actacaactc attcggatta acccctcatc cagtctctat 1261 aaatcattga tttcaggatt tgataaagaa aataaaaaag gttttcttat gagtttctta 1321 aaggagttgg cagaatatta tcaagctaaa gagagctgtg atgcggaaac tcagacaagc 1381 accacatttc ccagccaagt gtccctagcc gagaagtttc agcttattga cgctcaattt 1441 gcagatggtt ttcctcaccg ttccaagcta gaatctttag agacaaagtt aaacgagtat 1501 aagaaagaag tacaacacca gcttcaggta gaaatgtgtc ataagctgaa gtattttaga 1561 gaagctgaaa taacaaaagt taagatggaa gagagaagaa agtatgaaaa ggaattggct 1621 gagttccaga atgaatttga aagaacttgt caagcaaaaa atgaagccct catttctcag 1681 gagaagaatt ctctggaaag aattaaaaaa caccgagaga tggaatcaaa agaaatttat 1741 gctcaaagac agcttctact aaatgatata gctttgctta gaggtagaga agcagagctg 1801 aaggagcgaa tcgaaacttt tgaattgacc cagaagctcc aagaagaaaa aattaaaagt 1861 gaggctgagg cacttgagag acgggaacag aatctgaaga atattgaaga cacctatgat 1921 cagaaactca aaactgaact gctaaagtac caactggagc taaaggatga ctacatcact 1981 aggacaaata aactcctgga agaggaaagg aagaataaag aaaaaactat tcatttgcaa 2041 gaggagctca cagttattaa ttcaaaaaag gaagaactca gtaaatctgt taagcatatg 2101 aaagaagttg agctggaact ggagtctgtg aaagctcagt ttttagcaat atcaaaacaa 2161 aaccacttgc tgaatgaaaa ggtgagagag atgagtgatt actcacaact gaaagaagag 2221 aaagtggagc ttcaggcaca aaataaacta cttaaactgc aactggaaga gactaggaat 2281 gaaaacttac gactcctaga ccgcataact cagccacctc ctgagcttgt gatttttcag 2341 aaggaattac agaaaacaga aaaggctatg gaactggagc acaaggactt tgaaactcat 2401 aggcaggctc ttgagaaaca gctgcagagt gaaattgaaa attctgcaca gctaaggacc 2461 cagatagcag aatatgatgc ttctgtcaaa aggttaactg ttcaggtggc tgagttaaag 2521 tcacaactaa aacaaactca gatagcccta gagaatgaag tgtaccgaaa tccaaaacac 2581 tctctgattc attccctgag tggcctccta ttgagcggca agatggcacc ccacagtgaa 2641 gataaaagtg gggatttctt gaatgtaccc cttgaacaga acaaggttat agcgggtgca 2701 gttatgtcaa gggtcccacc ttatgtaaat acagcaacag aagctagttc cccagagtct 2761 gactttgagt ttatagccag tagtactaaa gccaaagtaa gagagctaga gcaagaggct 2821 gaacgcttgg aaaaggcatt cagaacttac tatcaaagag ctactcagaa tccttctacg 2881 agccctcagc cggcaaagag tcctccatct gtaaactcgg tggcagctct cagaagcata 2941 gcttccagtt ccatggacag acctgtttct gcagaggaca gagttgtctc ggaacagcct 3001 cttggggaca tgctgaagga agaaatgagt gatatgtcca aagcattcat gggcagtgtg 3061 gtttcacggc cacgcaggac ctcttcctcc acacgcctct cctctacacc ccatccaaaa 3121 tcgagaagaa gtcttgataa tgaaatgtat ctggaaggtc tcgggagatt acacatgact 3181 tcctccagcc ctcttcttga cagagtttct gcatcacctg ctgcttcccc tagtccttgt 3241 cctgagagaa cagcccaggc gtcacctgtt ccatctaggc acagcttctc tggtcttcca 3301 gagcaaaatg cctgtttata tcagagacaa actgaaacgc aagataaaag tgagctttca 3361 aatgtggaca agcaatcttt gaaagatgag aaatttgaac cacctttcag atggaacaag 3421 acagaacagt ttgaagcaga aggactccac cctgctggtg acatgcctgg aatagatttt 3481 gctgtggcta ctcagtcttc aagactcatt tcatacgact acccaagtgc tgttcagagc 3541 caaactggag aacaagacga acaggagcta tgggagcttc acatgaaaga acgcaggcag 3601 agagaagaac aaaggcataa tgaacggcaa gaagctttag agagggaaag aagagagctg 3661 ggaaaactag agcaggagcg gaggatgatt gaagagtcac tgaagatgga gatggaagaa 3721 gaattagaaa agagtgttca agaccagaaa gacaaatcag ctcattgtga aaatacttta 3781 gaaaaataca tgaaaattat ccagcagaga caggaagaat caaatgcaga taagagctcc 3841 aaaaaatcgg gcaaagaatg ctcactagtg gacatgatga tgcctagtga caaggatgaa 3901 agttctccag gattttctca tgaagaacca gatgacatgt gggagcaaaa gctcatttct 3961 gaagaggact tgtaaaaggg cgaattccag cacactggcg gccgttacta gtggatccat 4021 ccgcccctct ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg 4081 gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc 4141 ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa 4201 aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag 4261 acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg 4321 cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg 4381 ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa 4441 caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg 4501 gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca 4561 cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc atggtgagca 4621 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa 4681 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga 4741 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca 4801 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact 4861 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg 4921 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca 4981 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt 5041 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg 5101 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc 5161 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca 5221 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt 5281 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa agcggccgcg 5341 actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct 5401 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt 5461 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 5521 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt 5581 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat 5641 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga 5701 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca 5761 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct 5821 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc 5881 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag 5941 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca 6001 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg 6061 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga 6121 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa 6181 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg 6241 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg 6301 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc 6361 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca 6421 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt 6481 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga 6541 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg 6601 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg 6661 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt 6721 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg 6781 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat 6841 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat 6901 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg 6961 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg 7021 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc 7081 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc 7141 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg 7201 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg 7261 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca 7321 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac 7381 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga 7441 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga 7501 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac 7561 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt 7621 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc 7681 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc 7741 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc 7801 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat 7861 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt 7921 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct 7981 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc 8041 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc 8101 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc 8161 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc 8221 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg 8281 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata 8341 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta 8401 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc 8461 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg 8521 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt 8581 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt 8641 ggataaccgt attaccgcca tgcat //