LOCUS Exported 9208 bp ds-DNA circular SYN 15-JUN-2022 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Puro.Cre empty vector SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9208) AUTHORS Kumar MS, Erkeland SJ, Pester RE, Chen CY, Ebert MS, Sharp PA, Jacks T TITLE Suppression of non-small cell lung tumor development by the let-7 microRNA family. JOURNAL Proc Natl Acad Sci U S A. 2008 Feb 28. ():. PUBMED 18308936 REFERENCE 2 (bases 1 to 9208) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 15, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9208 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 192..211 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 263..283 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 284..1312 /codon_start=1 /product="site-specific recombinase" /label=Cre /note="Cre is a site-specific recombinase from bacteriophage P1. Recombination occurs at loxP sequences." /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED GD" misc_feature 1368..1956 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(1421..1441) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(1839..1850) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(1959..1975) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1960..1976) /label=pBluescriptKS /note="For pBluescript vector" LTR 2485..2665 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" rep_origin complement(2727..3315) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2807..2826) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3486..4346) /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 4109..4128 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4347..4451) /gene="bla" /label=AmpR promoter primer_bind complement(4526..4545) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 4717..5096 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 5098..5296 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5251..5271 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 5314..5494 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 5541..5666 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 6163..6396 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 6581..6625 /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" misc_feature 6923..7040 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 7109..8320 /label=UbC promoter /note="human ubiquitin C promoter" primer_bind 8157..8176 /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" CDS 8347..8946 /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(8347..8366) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 8843..8863 /label=Puro-F /note="Puromycin resistance gene, forward primer" primer_bind complement(9003..9026) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" ORIGIN 1 tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg 61 gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg 121 tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc 181 cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc 241 ctcatctccg ggacccggga tgcccaagaa gaagaggaag gtgtccaatt tactgaccgt 301 acaccaaaat ttgcctgcat taccggtcga tgcaacgagt gatgaggttc gcaagaacct 361 gatggacatg ttcagggatc gccaggcgtt ttctgagcat acctggaaaa tgcttctgtc 421 cgtttgccgg tcgtgggcgg catggtgcaa gttgaataac cggaaatggt ttcccgcaga 481 acctgaagat gttcgcgatt atcttctata tcttcaggcg cgcggtctgg cagtaaaaac 541 tatccagcaa catttgggcc agctaaacat gcttcatcgt cggtccgggc tgccacgacc 601 aagtgacagc aatgctgttt cactggttat gcggcggatc cgaaaagaaa acgttgatgc 661 cggtgaacgt gcaaaacagg ctctagcgtt cgaacgcact gatttcgacc aggttcgttc 721 actcatggaa aatagcgatc gctgccagga tatacgtaat ctggcatttc tggggattgc 781 ttataacacc ctgttacgta tagccgaaat tgccaggatc agggttaaag atatctcacg 841 tactgacggt gggagaatgt taatccatat tggcagaacg aaaacgctgg ttagcaccgc 901 aggtgtagag aaggcactta gcctgggggt aactaaactg gtcgagcgat ggatttccgt 961 ctctggtgta gctgatgatc cgaataacta cctgttttgc cgggtcagaa aaaatggtgt 1021 tgccgcgcca tctgccacca gccagctatc aactcgcgcc ctggaaggga tttttgaagc 1081 aactcatcga ttgatttacg gcgctaagga tgactctggt cagagatacc tggcctggtc 1141 tggacacagt gcccgtgtcg gagccgcgcg agatatggcc cgcgctggag tttcaatacc 1201 ggagatcatg caagctggtg gctggaccaa tgtaaatatt gtcatgaact atatccgtaa 1261 cctggatagt gaaacagggg caatggtgcg cctgctggaa gatggcgatt agccatgttt 1321 aagggttccg gttccactag gtacaattcg atatcaagct tatcgataat caacctctgg 1381 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat 1441 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt 1501 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca 1561 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg 1621 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg 1681 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca 1741 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca 1801 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc 1861 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc 1921 agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgatcga 1981 gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt 2041 gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct 2101 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg 2161 ggactggaag ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac 2221 cacacacaag gctacttccc tgattggcag aactacacac cagggccagg gatcagatat 2281 ccactgacct ttggatggtg ctacaagcta gtaccagttg agcaagagaa ggtagaagaa 2341 gccaatgaag gagagaacac ccgcttgtta caccctgtga gcctgcatgg gatggatgac 2401 ccggagagag aagtattaga gtggaggttt gacagccgcc tagcatttca tcacatggcc 2461 cgagagctgc atccggactg tactgggtct ctctggttag accagatctg agcctgggag 2521 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt 2581 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt 2641 tagtcagtgt ggaaaatctc tagcagcatg tgagcaaaag gccagcaaaa ggccaggaac 2701 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac 2761 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg 2821 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac 2881 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat 2941 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag 3001 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac 3061 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt 3121 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt 3181 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc 3241 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga 3301 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac 3361 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc 3421 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct 3481 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca 3541 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct 3601 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca 3661 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc 3721 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg 3781 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct 3841 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa 3901 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta 3961 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc 4021 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg 4081 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa 4141 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg 4201 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc 4261 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg 4321 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat 4381 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata 4441 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc 4501 ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt 4561 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta 4621 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg 4681 cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt 4741 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat 4801 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa 4861 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg 4921 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc 4981 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct 5041 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga 5101 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa 5161 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc 5221 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg 5281 aggtctatat aagcagcgcg ttttgcctgt actgggtctc tctggttaga ccagatctga 5341 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 5401 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 5461 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 5521 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 5581 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 5641 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 5701 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 5761 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 5821 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 5881 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 5941 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 6001 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 6061 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 6121 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 6181 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 6241 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 6301 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 6361 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 6421 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 6481 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 6541 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 6601 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 6661 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 6721 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 6781 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 6841 cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac 6901 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag 6961 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa 7021 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttag 7081 taccgggccc gctctagacg gttgatctgg cctccgcgcc gggttttggc gcctcccgcg 7141 ggcgcccccc tcctcacggc gagcgctgcc acgtcagacg aagggcgcag cgagcgtcct 7201 gatccttccg cccggacgct caggacagcg gcccgctgct cataagactc ggccttagaa 7261 ccccagtatc agcagaagga cattttagga cgggacttgg gtgactctag ggcactggtt 7321 ttctttccag agagcggaac aggcgaggaa aagtagtccc ttctcggcga ttctgcggag 7381 ggatctccgt ggggcggtga acgccgatga ttatataagg acgcgccggg tgtggcacag 7441 ctagttccgt cgcagccggg atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt 7501 cacttggtga gtagcgggct gctgggctgg ccggggcttt cgtggccgcc gggccgctcg 7561 gtgggacgga agcgtgtgga gagaccgcca agggctgtag tctgggtccg cgagcaaggt 7621 tgccctgaac tgggggttgg ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa 7681 tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg 7741 cggcaagaac ccaaggtctt gaggccttcg ctaatgcggg aaagctctta ttcgggtgag 7801 atgggctggg gcaccatctg gggaccctga cgtgaagttt gtcactgact ggagaactcg 7861 gtttgtcgtc tgttgcgggg gcggcagtta tggcggtgcc gttgggcagt gcacccgtac 7921 ctttgggagc gcgcgccctc gtcgtgtcgt gacgtcaccc gttctgttgg cttataatgc 7981 agggtggggc cacctgccgg taggtgtgcg gtaggctttt ctccgtcgca ggacgcaggg 8041 ttcgggccta gggtaggctc tcctgaatcg acaggcgccg gacctctggt gaggggaggg 8101 ataagtgagg cgtcagtttc tttggtcggt tttatgtacc tatcttctta agtagctgaa 8161 gctccggttt tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc 8221 accttttgaa atgtaatcat ttgggtcaat atgtaatttt cagtgttaga ctagtaaatt 8281 gtccgctaaa ttctggccgt ttttggcttt tttgttagac gaagctaacg cgctagcaaa 8341 gaattcatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg 8401 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat 8461 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg 8521 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg 8581 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg 8641 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc 8701 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt 8761 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc 8821 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc 8881 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt 8941 gcctgactcg agcggccgca ttctaccggg taggggaggc gcttttccca aggcagtctg 9001 gagcatgcgc tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc 9061 gcacacattc cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg 9121 cgccaccttc tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt 9181 gcaggacgtg acaaatggaa gtagcacg //