LOCUS Exported 9221 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS FUW SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9221) AUTHORS Lois C, Hong EJ, Pease S, Brown EJ, Baltimore D TITLE Germline transmission and tissue-specific expression of transgenes delivered by lentiviral vectors. JOURNAL Science. 2002 Feb 1. 295(5556):868-72. PUBMED 11786607 REFERENCE 2 (bases 1 to 9221) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9221 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(46..65) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 237..616 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 618..816 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 771..791 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 834..1014 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1061..1186 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1679..1912 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2097..2141 /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 2439..2556 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2621..3832 /label=UbC promoter /note="human ubiquitin C promoter" primer_bind 3669..3688 /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" misc_feature 3912..4500 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3965..3985) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4383..4394) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(4503..4519) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4504..4520) /label=pBluescriptKS /note="For pBluescript vector" LTR 5025..5205 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(5231..5248) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5237..5461 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 5507..5935 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5594..5613) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5804..5825 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5944..5964) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 5949..6278 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 6129..6264 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6191..6210 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 6326..6373 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 6392..6766 /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" polyA_signal 6896..7017 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6933..6952) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6987..7006 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(7066..7082) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7066..7082) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7079..7101) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7090..7106 /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(7114..7144) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7159..7180 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7297..7314) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7468..8056) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7548..7567) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8227..9087) /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 8850..8869 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9088..9192) /gene="bla" /label=AmpR promoter ORIGIN 1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga 61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg 121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct 181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca 241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata 301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga 361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt 421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt 481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca 541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt 601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt 661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca 721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg 781 cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc 841 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 901 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 961 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg 1021 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg 1081 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat 1141 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg 1201 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa 1261 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct 1321 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac 1381 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca 1441 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa 1501 aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga 1561 gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag 1621 tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag 1681 gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga 1741 cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc 1801 tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc 1861 tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt 1921 ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta 1981 ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta 2041 acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga 2101 atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa 2161 caaattggct gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa 2221 gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat 2281 cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag 2341 aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatcg gcactgcgtg 2401 cgccaattct gcagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt 2461 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa 2521 gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga 2581 gatccagttt ggttaattaa cccgtgtcgg ctccagatct ggcctccgcg ccgggttttg 2641 gcgcctcccg cgggcgcccc cctcctcacg gcgagcgctg ccacgtcaga cgaagggcgc 2701 agcgagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg ctcataagac 2761 tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt gggtgactct 2821 agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc ccttctcggc 2881 gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa ggacgcgccg 2941 ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt gtttgtggat 3001 cgctgtgatc gtcacttggt gagtagcggg ctgctgggct ggccggggct ttcgtggccg 3061 ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc caagggctgt agtctgggtc 3121 cgcgagcaag gttgccctga actgggggtt ggggggagcg cagcaaaatg gcggctgttc 3181 ccgagtcttg aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg 3241 gggcatggtg ggcggcaaga acccaaggtc ttgaggcctt cgctaatgcg ggaaagctct 3301 tattcgggtg agatgggctg gggcaccatc tggggaccct gacgtgaagt ttgtcactga 3361 ctggagaact cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca 3421 gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt 3481 ggcttataat gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg 3541 caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg 3601 gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct 3661 taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga 3721 agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta 3781 gactagtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag acgaagcttg 3841 ggctgcaggt cgactctaga ggatccagtt aacctcgagg gcgcgccgaa ttcgatatca 3901 agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta 3961 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta 4021 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt 4081 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg 4141 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt 4201 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag 4261 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc 4321 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc 4381 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc 4441 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc 4501 atcgataccg tcgacctcga gacctagaaa aacatggagc aatcacaagt agcaatacag 4561 cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc 4621 cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc 4681 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagata 4741 tccttgatct gtggatctac cacacacaag gctacttccc tgattggcag aactacacac 4801 cagggccagg gatcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg 4861 agcaagagaa ggtagaagaa gccaatgaag gagagaacac ccgcttgtta caccctgtga 4921 gcctgcatgg gatggatgac ccggagagag aagtattaga gtggaggttt gacagccgcc 4981 tagcatttca tcacatggcc cgagagctgc atccggactg tactgggtct ctctggttag 5041 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat 5101 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact 5161 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagggcc cgtttaaacc 5221 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc 5281 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa 5341 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac 5401 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg 5461 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc 5521 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc 5581 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt 5641 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac 5701 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag 5761 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa 5821 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg 5881 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc 5941 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta 6001 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag 6061 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa 6121 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 6181 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt 6241 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat 6301 ccattttcgg atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat 6361 agtataatac gacaaggtga ggaactaaac catggccaag ttgaccagtg ccgttccggt 6421 gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc 6481 ccgggacttc gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat 6541 cagcgcggtc caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg 6601 cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc 6661 cgggccggcc atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga 6721 cccggccggc aactgcgtgc acttcgtggc cgaggagcag gactgacacg tgctacgaga 6781 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc 6841 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt 6901 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 6961 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 7021 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc 7081 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 7141 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 7201 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 7261 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 7321 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 7381 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 7441 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 7501 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 7561 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 7621 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 7681 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 7741 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 7801 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 7861 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg 7921 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 7981 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 8041 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 8101 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 8161 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 8221 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 8281 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 8341 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 8401 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 8461 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 8521 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 8581 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 8641 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 8701 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 8761 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 8821 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 8881 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 8941 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 9001 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 9061 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 9121 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 9181 aggggttccg cgcacatttc cccgaaaagt gccacctgac g //