LOCUS Exported 7490 bp ds-DNA circular SYN 14-SEP-2021 DEFINITION Expresses human Peg10 cargoRNA encoding Cre recombinase. ACCESSION . VERSION . KEYWORDS pCMV-Hs.cargo(Cre) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7490) AUTHORS Segel M, Lash B, Song J, Ladha A, Liu CC, Jin X, Mekhedov SL, Macrae RK, Koonin EV, Zhang F TITLE Mammalian retrovirus-like protein PEG10 packages its own mRNA and can be pseudotyped for mRNA delivery. JOURNAL Science. 2021 Aug 20;373(6557):882-889. doi: 10.1126/science.abg6155. PUBMED 34413232 REFERENCE 2 (bases 1 to 7490) AUTHORS . TITLE Direct Submission JOURNAL Exported Sep 14, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7490 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 381..584 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 535..555 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 629..648 /label=T7 /note="T7 promoter, forward primer" promoter 629..647 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 1156..1165 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1168..1188 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1192..2217 /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="NLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLLS VCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLPR PSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFLG IAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVERW ISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQRY LAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLEDG D" primer_bind complement(2847..2864) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2853..3077 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3123..3551 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3210..3229) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3420..3441 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3560..3580) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3565..3894 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3745..3880 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3807..3826 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3961..4755 /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 GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4015..4034) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4625..4644 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4929..5050 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4966..4985) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5020..5039 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5099..5115) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5099..5115) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5112..5134) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5123..5139 /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)." promoter complement(5147..5177) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5192..5213 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5330..5347) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5501..6089) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5581..5600) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6260..7120) /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 6883..6902 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7121..7225) /gene="bla" /label=AmpR promoter primer_bind complement(7300..7319) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag 601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg 661 gctagcgttt aaaaccctcc tcggtgcaac ctatataagg ctcacagtct gcgctcctgg 721 tacacgcgct tcaacttcgg ttggtgtgtg tcgaagaaac ctgactgcgc cctgaggaga 781 acagcggaga aggtccaccg agcctggcga aaggtccgct gagcgggctg tcgtccggag 841 ccactccggg ctgcggagca cccagtggag accgcgcctg gctcaggtgt gggaccccat 901 ccttcctgtc ttcgcagagg agtcctcgcg tgaaataagc gggttttgaa aacaaaaaaa 961 agaaggagtg gaagaggggg ccaggatcca ggcctccatc cccacagaag tgaagctaca 1021 gctgggaggt ctcctcccac cccaaccgtc accctgggtc ccgactgccc acctcctcct 1081 cctccccctc cccccaacaa caacaacaac aacaactcca agcacaccgg ccataagagt 1141 gcgtgtgtcc ccaacgccac catgggccca aagaagaaga gaaaggtttc gaatttactg 1201 accgtacacc aaaatttgcc tgcattaccg gtcgatgcaa cgagtgatga ggttcgcaag 1261 aacctgatgg acatgttcag ggatcgccag gcgttttctg agcatacctg gaaaatgctt 1321 ctgtccgttt gccggtcgtg ggcggcatgg tgcaagttga ataaccggaa atggtttccc 1381 gcagaacctg aagatgttcg cgattatctt ctatatcttc aggcgcgcgg tctggcagta 1441 aaaactatcc agcaacattt gggccagcta aacatgcttc atcgtcggtc cgggctgcca 1501 cgaccaagtg acagcaatgc tgtttcactg gttatgcggc ggatccgaaa agaaaacgtt 1561 gatgccggtg aacgtgcaaa acaggctcta gcgttcgaac gcactgattt cgaccaggtt 1621 cgttcactca tggaaaatag cgatcgctgc caggatatac gtaatctggc atttctgggg 1681 attgcttata acaccctgtt acgtatagcc gaaattgcca ggatcagggt taaagatatc 1741 tcacgtactg acggtgggag aatgttaatc catattggca gaacgaaaac gctggttagc 1801 accgcaggtg tagagaaggc acttagcctg ggggtaacta aactggtcga gcgatggatt 1861 tccgtctctg gtgtagctga tgatccgaat aactacctgt tttgccgggt cagaaaaaat 1921 ggtgttgccg cgccatctgc caccagccag ctatcaactc gcgccctgga agggattttt 1981 gaagcaactc atcgattgat ttacggcgct aaggatgact ctggtcagag atacctggcc 2041 tggtctggac acagtgcccg tgtcggagcc gcgcgagata tggcccgcgc tggagtttca 2101 ataccggaga tcatgcaagc tggtggctgg accaatgtaa atattgtcat gaactatatc 2161 cgtaacctgg atagtgaaac aggggcaatg gtgcgcctgc tggaagatgg cgattaaata 2221 cctgtcatgt ccttcaggat ctctgccctc aaaatttatt cctgttcagc ttctcaatca 2281 gtgactgtgt gctaaatttt aggctactgt atcttcaggc cacctgaggc acatcctctc 2341 tgaaacggct atggaaggtt agggccactc tggactggca cacatcctaa agcaccaaaa 2401 gaccttcaac attttctgag agcaacagag tatttgccaa taaatgatct ctcatttttc 2461 caccttgact gccaatctaa ctaaaataat taataagttt actttccagc cagtcctgga 2521 agtctgggtt ttacctgcca aaacctccat caccatctaa attataggct gccaaatttg 2581 ctgtttaaca tttacagaga agctgataca aacgcaggaa atgctgattt ctttatggag 2641 ggggagacga ggaggaggag gacatgactt ttcttgcggt ttcggtaccc tctttttaaa 2701 tcactggagg actgaggcct tattaaggaa gccttaagct tggtaccgag ctcggatcca 2761 ctagtccagt gtggtggaat tctgcagata tccagcacag tggcggccgc tcgagtctag 2821 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt 2881 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc 2941 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg 3001 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga 3061 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc 3121 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac 3181 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc 3241 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt 3301 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg 3361 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag 3421 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt 3481 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt 3541 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc 3601 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag 3661 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc 3721 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 3781 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct 3841 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc 3901 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc 3961 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc 4021 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca 4081 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg 4141 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg 4201 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag 4261 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg 4321 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc 4381 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa 4441 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac 4501 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat 4561 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac 4621 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc 4681 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt 4741 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc 4801 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg 4861 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg 4921 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 4981 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 5041 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt 5101 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 5161 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 5221 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 5281 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 5341 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 5401 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 5461 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 5521 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 5581 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 5641 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 5701 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 5761 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 5821 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 5881 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 5941 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 6001 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 6061 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 6121 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 6181 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 6241 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 6301 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 6361 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 6421 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 6481 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 6541 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 6601 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 6661 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 6721 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 6781 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 6841 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 6901 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 6961 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 7021 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 7081 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 7141 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 7201 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg 7261 gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc 7321 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag 7381 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag 7441 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt //