LOCUS Exported 5645 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pLenti X2 Zeo/pTER shGFP #2 (790-1) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5645) TITLE shMDC1 plasmids REFERENCE 2 (bases 1 to 5645) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5645 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 72..660 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 561..580 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 814..831 /label=L4440 /note="L4440 vector, forward primer" protein_bind 948..969 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 984..1014 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1022..1038 /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 1027..1049 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 1046..1062 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 1046..1062 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 1081..1101 /label=T3 /note="T3 promoter, forward primer" promoter 1083..1101 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" LTR 1357..1537 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1581..1706 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 2199..2432 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2616..2660 /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 2921..3038 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" misc_feature 3092..3680 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3145..3165) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3563..3574) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(3731..3751) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3736..4066 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3917..4052 /label=SV40 ori /note="SV40 origin of replication" promoter 4078..4125 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 4144..4518 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" protein_bind 4654..4678 /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind 4810..4828 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind complement(4874..4893) /label=H1 /note="Human H1 promoter, forward primer" protein_bind complement(5070..5094) /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" LTR 5169..5349 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" polyA_signal 5421..5542 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5458..5477) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5512..5531 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" ORIGIN 1 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc 61 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa 121 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag 181 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta 241 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta 301 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag 361 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg 421 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg 481 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag 541 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc 601 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa 661 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg 721 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct 781 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa 841 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg 901 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag 961 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga 1021 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc 1081 gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg tagtcttatg 1141 caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc ttacaaggag 1201 agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt gccttattag 1261 gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg cattgcagag 1321 atattgtatt taagtgccta gctcgataca ataaacgggt ctctctggtt agaccagatc 1381 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg 1441 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc 1501 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacctga 1561 aagcgaaagg gaaaccagag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 1621 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 1681 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 1741 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 1801 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 1861 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1921 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1981 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 2041 aagcggccgc tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa 2101 ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag 2161 agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc 2221 ttgggagcag caggaagcac tatgggcgca gcctcaatga cgctgacggt acaggccaga 2281 caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa 2341 cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct 2401 gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc 2461 atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt 2521 ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac 2581 actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat 2641 tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa 2701 aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt 2761 ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa 2821 ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag 2881 acagatccat tcgattagtg aacggatctc gacggttaac ttttaaaaga aaagggggga 2941 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta 3001 aagaattaca aaaacaaatt acaaaaattc aaaattttat tccagtgtgg tggaattctg 3061 cagatatcca gcacagtggc ggccgctcga caatcaacct ctggattaca aaatttgtga 3121 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt 3181 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa 3241 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt 3301 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct 3361 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg 3421 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc 3481 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg 3541 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct 3601 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc 3661 cctttgggcc gcctccccgc ctggaattct gcagatatcc ggttagtaat gagtttggaa 3721 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccaggcagg 3781 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg 3841 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc 3901 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca 3961 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt 4021 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tccccctgtt 4081 gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa 4141 accatggcca agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg 4201 gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc 4261 ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg 4321 gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg 4381 gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag 4441 cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg 4501 gccgaggagc aggactgaca cgtgctacga gatttaaatg gtacctttaa gaccaatgac 4561 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct 4621 agactcgagc ggccgccact gtgctggata tcaaccactt tgtacaagaa agctgggtct 4681 agatatctcg agtgcggccg cgaattccac cacactggac tagtggatcc gagctcggta 4741 ccaagctttt ccaaaaagct ggagtacaac tacaactctc ttgaagttgt agttgtactc 4801 cagcggggat ctctatcact gatagggaac ttataagatt cccaaatcca aagacatttc 4861 acgtttatgg tgatttccca gaacacatag cgacatgcaa atattgcagg gcgccactcc 4921 cctgtccctc acagccatct tcctgccagg gcgcacgcgc gctgggtgtt cccgcctagt 4981 gacactgggc cgcgattcct tggagcgggt tgatgacgtc agcgttcgaa ttcggtaccg 5041 gatccagtcg actgaattgg ttcctttaaa gcctgctttt ttgtacaaac ttgttgatat 5101 ctgcagaatt ccaccacact ggactagctc actcccaacg aagacaagat ctgctttttg 5161 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctgcctag 5221 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc 5281 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa 5341 tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat 5401 gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca 5461 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt 5521 ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc 5581 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga 5641 ggccg //