LOCUS Exported 8547 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Express copGFP and iCre under CB promoter. ACCESSION . VERSION . KEYWORDS pCDH-CB-copGFP-T2A-iCre SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8547) TITLE lenti vector JOURNAL Unpublished REFERENCE 2 (bases 1 to 8547) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8547 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 234..414 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 458..583 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1076..1309 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1493..1537 /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 1798..1915 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" intron 2567..2663 /label=SV40 intron /note="modified SV40 intron with splice donor and acceptor sites" CDS 2765..3427 /codon_start=1 /product="green fluorescent protein 2 from Pontellina plumata, also known as ppluGFP2 (Shagin et al., 2004)" /label=CopGFP /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA" CDS 3512..3565 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" CDS 3572..3592 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" misc_feature 4642..5230 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4695..4715) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5113..5124) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 5304..5537 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 5609..5730 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5646..5665) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5700..5719 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 5749..5884 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5811..5830 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(5922..5938) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5922..5938) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5935..5957) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5946..5962 /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(5970..6000) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6015..6036 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6153..6170) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6324..6912) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6404..6423) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7083..7943) /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 7706..7725 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7944..8048) /gene="bla" /label=AmpR promoter primer_bind 8116..8134 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(8172..8194) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 8294..8313 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 8507..8529 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 8521..8538 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 8522..8538 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" ORIGIN 1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca 61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta 121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga 181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc 241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg 421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct 481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt 541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag 601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt 661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt 721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg 781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag 841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag 901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg 961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag 1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag 1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc 1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga 1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc 1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg 1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata 1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa 1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga 1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa 1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat 1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt 1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg 1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggttaacttt 1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc 1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga 1921 ttaattaagg tacctctggt cgttacataa cttacggtaa atggcccgcc tggctgaccg 1981 cccaacgacc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 2041 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 2101 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 2161 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctact 2221 cgaggccacg ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta 2281 tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg gggggggggc 2341 gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg 2401 cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc 2461 ggcggcccta taaaaagcga agcgcgcggc gggcgggagc gggatcagcc accgcggtgg 2521 cggccctaga gtcgatcgag gaactgaaaa accagaaagt taactggtaa gtttagtctt 2581 tttgtctttt atttcaggtc cccgatccgg tggtggtgca aatcaaagaa ctgctcctca 2641 gtggatgttg cctttacttc taggcctgta cggaagtgtt acttctgctc taaaagctgc 2701 ggaattgtac ccgcggccga tccaccggtc gggatccacc atggagagcg acgagagcgg 2761 cctgcccgcc atggagatcg agtgccgcat caccggcacc ctgaacggcg tggagttcga 2821 gctggtgggc ggcggagagg gcacccccaa gcagggccgc atgaccaaca agatgaagag 2881 caccaaaggc gccctgacct tcagccccta cctgctgagc cacgtgatgg gctacggctt 2941 ctaccacttc ggcacctacc ccagcggcta cgagaacccc ttcctgcacg ccatcaacaa 3001 cggcggctac accaacaccc gcatcgagaa gtacgaggac ggcggcgtgc tgcacgtgag 3061 cttcagctac cgctacgagg ccggccgcgt gatcggcgac ttcaaggtgg tgggcaccgg 3121 cttccccgag gacagcgtga tcttcaccga caagatcatc cgcagcaacg ccaccgtgga 3181 gcacctgcac cccatgggcg ataacgtgct ggtgggcagc ttcgcccgca ccttcagcct 3241 gcgcgacggc ggctactaca gcttcgtggt ggacagccac atgcacttca agagcgccat 3301 ccaccccagc atcctgcaga acgggggccc catgttcgcc ttccgccgcg tggaggagct 3361 gcacagcaac accgagctgg gcatcgtgga gtaccagcac gccttcaaga cccccattgc 3421 cttcgccaga tcccgcgctc agtcgtccaa ttctgccgtg gacggcaccg ccggacccgg 3481 ctccaccgga tctcgcaagc ttggcagtgg agagggcaga ggaagtctgc taacatgcgg 3541 tgacgtcgag gagaatcctg gcccaatggt gcccaagaag aagaggaaag tctccaacct 3601 gctgactgtg caccaaaacc tgcctgccct ccctgtggat gccacctctg atgaagtcag 3661 gaagaacctg atggacatgt tcagggacag gcaggccttc tctgaacaca cctggaagat 3721 gctcctgtct gtgtgcagat cctgggctgc ctggtgcaag ctgaacaaca ggaaatggtt 3781 ccctgctgaa cctgaggatg tgagggacta cctcctgtac ctgcaagcca gaggcctggc 3841 tgtgaagacc atccaacagc acctgggcca gctcaacatg ctgcacagga gatctggcct 3901 gcctcgccct tctgactcca atgctgtgtc cctggtgatg aggagaatca gaaaggagaa 3961 tgtggatgct ggggagagag ccaagcaggc cctggccttt gaacgcactg actttgacca 4021 agtcagatcc ctgatggaga actctgacag atgccaggac atcaggaacc tggccttcct 4081 gggcattgcc tacaacaccc tgctgcgcat tgccgaaatt gccagaatca gagtgaagga 4141 catctcccgc accgatggtg ggagaatgct gatccacatt ggcaggacca agaccctggt 4201 gtccacagct ggtgtggaga aggccctgtc cctgggggtt accaagctgg tggagagatg 4261 gatctctgtg tctggtgtgg ctgatgaccc caacaactac ctgttctgcc gggtcagaaa 4321 gaatggtgtg gctgcccctt ctgccacctc ccaactgtcc acccgggccc tggaagggat 4381 ctttgaggcc acccaccgcc tgatctatgg tgccaaggat gactctgggc agagatacct 4441 ggcctggtct ggccactctg ccagagtggg tgctgccagg gacatggcca gggctggtgt 4501 gtccatccct gaaatcatgc aggctggtgg ctggaccaat gtgaacattg tgatgaacta 4561 catcagaaac ctggactctg agactggggc catggtgagg ctgctcgagg atggggactg 4621 agcggccgcg aattcgtcga caatcaacct ctggattaca aaatttgtga aagattgact 4681 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg 4741 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg 4801 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg 4861 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg 4921 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc 4981 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca 5041 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc 5101 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct 5161 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc 5221 gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca 5281 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa aataagatct 5341 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg 5401 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt 5461 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt 5521 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc 5581 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa 5641 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 5701 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta 5761 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca 5821 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga 5881 ggcctagact tttgcagaga cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg 5941 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 6001 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 6061 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 6121 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 6181 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 6241 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 6301 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 6361 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 6421 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 6481 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 6541 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 6601 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 6661 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 6721 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc 6781 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 6841 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 6901 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 6961 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 7021 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 7081 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 7141 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 7201 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 7261 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 7321 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 7381 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 7441 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 7501 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 7561 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 7621 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 7681 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 7741 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 7801 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 7861 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 7921 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 7981 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 8041 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg 8101 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat 8161 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg 8221 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc 8281 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa 8341 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg 8401 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa 8461 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt 8521 tgtaaaacga cggccagtgc caagctg //