LOCUS Exported 7725 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Express iCre under CB promoter. ACCESSION . VERSION . KEYWORDS pCDH-CB-iCre SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7725) TITLE lenti vector JOURNAL Unpublished REFERENCE 2 (bases 1 to 7725) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7725 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 6..232 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 233..413 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 460..585 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1078..1311 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1496..1540 /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" intron 2575..2671 /label=SV40 intron /note="modified SV40 intron with splice donor and acceptor sites" CDS 2755..2775 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" misc_feature 3825..4413 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3878..3898) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4296..4307) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 4487..4720 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 4792..4913 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4829..4848) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4883..4902 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 4932..5067 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4994..5013 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(5100..5116) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5100..5116) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5113..5135) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5124..5140 /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(5148..5178) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5193..5214 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5331..5348) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5502..6090) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5582..5601) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6261..7121) /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 6884..6903 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7122..7226) /gene="bla" /label=AmpR promoter primer_bind 7294..7312 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(7350..7372) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 7472..7491 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7685..7707 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 7699..7716 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 7700..7716 /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 cgatacataa acgggtctct 241 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa 301 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc 361 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc 421 ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg 481 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt 541 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg 601 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa 661 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg 721 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca 781 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa 841 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa 901 agtaagacca ccgcacagca agcggccact gatcttcaga cctggaggag gagatatgag 961 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt 1021 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg 1081 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac 1141 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct 1201 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct 1261 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg 1321 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa 1381 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa 1441 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa 1501 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac 1561 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag 1621 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc 1681 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga 1741 aggtggagag agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg 1801 gttaactttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga 1861 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aataccttaa 1921 ttaatcgatt aattaaggta cctctggtcg ttacataact tacggtaaat ggcccgcctg 1981 gctgaccgcc caacgacccc gcccattgac gtcaataatg acgtatgttc ccatagtaac 2041 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt 2101 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa 2161 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta 2221 catctactcg aggccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca 2281 attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg gggggggggg 2341 gggggggcgc gcgccggggg gggggggggg gggggggggg gggggggggg gggggcggag 2401 aggtgcggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg 2461 gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcgggagcgg gatcagccac 2521 cgcggtggcg gccctagagt cgatcgagga actgaaaaac cagaaagtta actggtaagt 2581 ttagtctttt tgtcttttat ttcaggtccc cgatccggtg gtggtgcaaa tcaaagaact 2641 gctcctcagt ggatgttgcc tttacttcta ggcctgtacg gaagtgttac ttctgctcta 2701 aaagctgcgg aattgtaccc gcggccgatc caccggtcgg gatccaccat ggtgcccaag 2761 aagaagagga aagtctccaa cctgctgact gtgcaccaaa acctgcctgc cctccctgtg 2821 gatgccacct ctgatgaagt caggaagaac ctgatggaca tgttcaggga caggcaggcc 2881 ttctctgaac acacctggaa gatgctcctg tctgtgtgca gatcctgggc tgcctggtgc 2941 aagctgaaca acaggaaatg gttccctgct gaacctgagg atgtgaggga ctacctcctg 3001 tacctgcaag ccagaggcct ggctgtgaag accatccaac agcacctggg ccagctcaac 3061 atgctgcaca ggagatctgg cctgcctcgc ccttctgact ccaatgctgt gtccctggtg 3121 atgaggagaa tcagaaagga gaatgtggat gctggggaga gagccaagca ggccctggcc 3181 tttgaacgca ctgactttga ccaagtcaga tccctgatgg agaactctga cagatgccag 3241 gacatcagga acctggcctt cctgggcatt gcctacaaca ccctgctgcg cattgccgaa 3301 attgccagaa tcagagtgaa ggacatctcc cgcaccgatg gtgggagaat gctgatccac 3361 attggcagga ccaagaccct ggtgtccaca gctggtgtgg agaaggccct gtccctgggg 3421 gttaccaagc tggtggagag atggatctct gtgtctggtg tggctgatga ccccaacaac 3481 tacctgttct gccgggtcag aaagaatggt gtggctgccc cttctgccac ctcccaactg 3541 tccacccggg ccctggaagg gatctttgag gccacccacc gcctgatcta tggtgccaag 3601 gatgactctg ggcagagata cctggcctgg tctggccact ctgccagagt gggtgctgcc 3661 agggacatgg ccagggctgg tgtgtccatc cctgaaatca tgcaggctgg tggctggacc 3721 aatgtgaaca ttgtgatgaa ctacatcaga aacctggact ctgagactgg ggccatggtg 3781 aggctgctcg aggatgggga ctgagcggcc gcgaattcgt cgacaatcaa cctctggatt 3841 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg 3901 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct 3961 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc 4021 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca 4081 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac 4141 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt 4201 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct 4261 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc 4321 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga 4381 cgagtcggat ctccctttgg gccgcctccc cgcctggtac ctttaagacc aatgacttac 4441 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt 4501 cactcccaac gaagataaga tctgcttttt gcttgtactg ggtctctctg gttagaccag 4561 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc 4621 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga 4681 tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt 4741 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt 4801 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca 4861 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc 4921 tggctctagc tatcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct 4981 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga 5041 agtagtgagg aggctttttt ggaggcctag acttttgcag agaccaaatt cgtaatcatg 5101 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc 5161 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg 5221 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc 5281 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact 5341 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta 5401 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag 5461 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc 5521 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta 5581 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg 5641 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc 5701 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac 5761 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac 5821 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg 5881 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga 5941 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt 6001 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag 6061 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 6121 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 6181 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat 6241 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc 6301 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg 6361 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct 6421 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca 6481 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg 6541 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg 6601 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc 6661 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag 6721 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg 6781 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag 6841 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat 6901 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg 6961 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca 7021 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca 7081 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat 7141 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag 7201 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa 7261 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt 7321 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc 7381 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt 7441 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg 7501 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc 7561 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta 7621 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg 7681 ttttcccagt cacgacgttg taaaacgacg gccagtgcca agctg //