LOCUS Exported 7084 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pLKO.1 Sox2 HM a SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7084) AUTHORS Bass AJ, Watanabe H, Mermel CH, Yu S, Perner S, Verhaak RG, Kim SY, Wardwell L, Tamayo P, Gat-Viks I, Ramos AH, Woo MS, Weir BA, Getz G, Beroukhim R, O'Kelly M, Dutt A, Rozenblatt-Rosen O, Dziunycz P, Komisarof J, Chirieac LR, Lafargue CJ, Scheble V, Wilbertz T, Ma C, Rao S, Nakagawa H, Stairs DB, Lin L, Giordano TJ, Wagner P, Minna JD, Gazdar AF, Zhu CQ, Brose MS, Cecconello I, Jr UR, Marie SK, Dahl O, Shivdasani RA, Tsao MS, Rubin MA, Wong KK, Regev A, Hahn WC, Beer DG, Rustgi AK, Meyerson M TITLE SOX2 is an amplified lineage-survival oncogene in lung and esophageal squamous cell carcinomas. JOURNAL Nat Genet. 2009 Nov . 41(11):1238-42. PUBMED 19801978 REFERENCE 2 (bases 1 to 7084) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7084 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 128..147 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 381..398 /label=L4440 /note="L4440 vector, forward primer" protein_bind 515..536 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 551..581 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 589..605 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 594..616 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 613..629 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 613..629 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 648..668 /label=T3 /note="T3 promoter, forward primer" promoter 650..668 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 696..922 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 923..1103 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1150..1275 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1768..2001 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2186..2230 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" promoter 2528..2768 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 2528..2548 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 2699..2718 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_feature 2873..2990 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 3039..3549 /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" primer_bind 3468..3486 /label=hPGK-F /note="Human PGK promoter, forward primer" CDS 3571..4170 /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(3571..3590) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 4067..4087 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 4298..4531 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 4603..4724 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4640..4659) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4694..4713 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 4764..4899 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4826..4845 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(4919..4938) /label=T7 /note="T7 promoter, forward primer" promoter complement(4920..4938) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4948..4965) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4948..4964) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4957..4979) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 5106..5561 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5193..5212) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5403..5424 /label=F1ori-F /note="F1 origin, forward primer" promoter 5587..5691 /gene="bla" /label=AmpR promoter CDS 5692..6552 /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 complement(5910..5929) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6723..227 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga 61 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc 121 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg 181 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg 241 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt 301 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc 361 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac 421 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc 481 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg 541 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat 601 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc 661 actaaaggga acaaaagctg gagctgcaag cttaatgtag tcttatgcaa tactcttgta 721 gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga aaaagcaccg 781 tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa ggcaacagac 841 gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata ttgtatttaa 901 gtgcctagct cgatacataa acgggtctct ctggttagac cagatctgag cctgggagct 961 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca 1021 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta 1081 gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa 1141 ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag 1201 gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat 1261 gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg 1321 ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag 1381 ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata 1441 ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat 1501 acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct 1561 ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct 1621 gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata 1681 taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt 1741 gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc 1801 aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc 1861 tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt 1921 gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata 1981 cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac 2041 tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac 2101 gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat 2161 tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg 2221 ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa aattattcat 2281 aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa 2341 tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg 2401 acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag acagatccat 2461 tcgattagtg aacggatctc gacggtatcg atcacgagac tagcctcgag cggccgcccc 2521 cttcaccgag ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt 2581 tagagagata attggaatta atttgactgt aaacacaaag atattagtac aaaatacgtg 2641 acgtagaaag taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga 2701 ctatcatatg cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg 2761 gaaaggacga aacaccggcg agataaacat ggcaatcaac tcgagttgat tgccatgttt 2821 atctcgtttt tgaattctcg acctcgagac aaatggcagt attcatccac aattttaaaa 2881 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag 2941 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt 3001 acagggacag cagagatcca ctttggccgc ggctcgaggg ggttggggtt gcgccttttc 3061 caaggcagcc ctgggtttgc gcagggacgc ggctgctctg ggcgtggttc cgggaaacgc 3121 agcggcgccg accctgggtc tcgcacattc ttcacgtccg ttcgcagcgt cacccggatc 3181 ttcgccgcta cccttgtggg ccccccggcg acgcttcctg ctccgcccct aagtcgggaa 3241 ggttccttgc ggttcgcggc gtgccggacg tgacaaacgg aagccgcacg tctcactagt 3301 accctcgcag acggacagcg ccagggagca atggcagcgc gccgaccgcg atgggctgtg 3361 gccaatagcg gctgctcagc agggcgcgcc gagagcagcg gccgggaagg ggcggtgcgg 3421 gaggcggggt gtggggcggt agtgtgggcc ctgttcctgc ccgcgcggtg ttccgcattc 3481 tgcaagcctc cggagcgcac gtcggcagtc ggctccctcg ttgaccgaat caccgacctc 3541 tctccccagg gggatccacc ggagcttacc atgaccgagt acaagcccac ggtgcgcctc 3601 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac 3661 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa 3721 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc 3781 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag 3841 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa 3901 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc 3961 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg 4021 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc 4081 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc 4141 tggtgcatga cccgcaagcc cggtgcctga cgcccgcccc acgacccgca gcgcccgacc 4201 gaaaggagcg cacgacccca tgcatcggta cctttaagac caatgactta caaggcagct 4261 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa 4321 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc 4381 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga 4441 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga 4501 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag 4561 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct 4621 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca 4681 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag 4741 ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc 4801 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg 4861 agctattcca gaagtagtga ggaggctttt ttggaggcct agggacgtac ccaattcgcc 4921 ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga 4981 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg 5041 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga 5101 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt 5161 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct 5221 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg 5281 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag 5341 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa 5401 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga 5461 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa 5521 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga 5581 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc 5641 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt 5701 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct 5761 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt 5821 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt 5881 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac 5941 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac 6001 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct 6061 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg 6121 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg 6181 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca 6241 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa 6301 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt 6361 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc 6421 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg 6481 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt 6541 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt 6601 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc 6661 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct 6721 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta 6781 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc 6841 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac 6901 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct 6961 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat 7021 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg 7081 acct //