LOCUS Exported 7448 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Luc-pSUPER-Retro-GFP/Neo SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7448) AUTHORS Li L, Zhang L, Zhang X, Yan Q, Minamishima YA, Olumi AF, Mao M, Bartz S, Kaelin WG TITLE Hypoxia-inducible factor linked to differential kidney cancer risk seen with type 2A and type 2B VHL mutations. JOURNAL Mol Cell Biol. 2007 Aug . 27(15):5381-92. PUBMED 17526729 REFERENCE 2 (bases 1 to 7448) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7448 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 579..920 /label=MESV Psi /note="packaging signal of murine embryonic stem cell virus" CDS 987..1403 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1333..1355 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1371..1387 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" primer_bind 1421..1437 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 1422..1438 /label=KS primer /note="common sequencing primer, one of multiple similar variants" promoter complement(1507..1721) /label=H1 promoter /note="human H1 RNA promoter" primer_bind complement(1567..1586) /label=H1 /note="Human H1 promoter, forward primer" primer_bind complement(1767..1790) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 2168..2187 /label=mPGK-F /note="Mouse PGK promoter, forward primer" regulatory 2264..2273 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2270..2986 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 2270..2986 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2315..2336) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2576..2595) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2923..2944 /label=EGFP-C /note="EGFP, forward primer" CDS 3050..3844 /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(3104..3123) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3714..3733 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3908..3956 /note="synthetic polyadenylation signal" protein_bind 4517..4533 /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(4541..4571) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4586..4607 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4724..4741) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4895..5483) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4975..4994) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5654..6514) /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 6277..6296 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6515..6619) /gene="bla" /label=AmpR promoter primer_bind 6687..6705 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(6743..6765) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 6865..6884 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7078..7100 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 7228..7247 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" LTR 7447..515 /label=5' LTR /note="5' long terminal repeat from murine embryonic stem cell virus" ORIGIN 1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat 61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca 121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga 181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt 241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc 301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc 361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata 421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca 481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga 541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg 601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt 661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa 721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga 781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag 841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa 901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg 961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct 1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga 1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag 1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc 1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc 1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg 1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag 1381 ccctcactcc ttctctaggc gccggaatta gatcgatctc tcgaggtcga cggtatcgat 1441 aagcttaaaa acgtacgcgg aatacttcga tctcttgaat cgaagtattc cgcgtacggg 1501 ggatctgtgg tctcatacag aacttataag attcccaaat ccaaagacat ttcacgttta 1561 tggtgatttc ccagaacaca tagcgacatg caaatattgc agggcgccac tcccctgtcc 1621 ctcacagcca tcttcctgcc agggcgcacg cgcgctgggt gttcccgcct agtgacactg 1681 ggcccgcgat tccttggagc gggttgatga cgtcagcgtt cgaattctac cgggtagggg 1741 aggcgctttt cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc 1801 gctacacaag tggcctctgg cctcgcacac attccacatc caccggccgg taggcgccaa 1861 ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct agtcaggaag 1921 ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc 1981 actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg cctttggggc 2041 agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg aaggggtggg 2101 tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga 2161 ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca 2221 tctccgggcc tttcgacctg cagcccaagc tagcttaccg gtcgccacca tggtgagcaa 2281 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa 2341 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac 2401 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac 2461 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt 2521 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga 2581 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat 2641 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta 2701 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt 2761 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca 2821 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac 2881 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt 2941 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtccg gactcagatc 3001 agcttgattc ttctgacaca acagtctcga acttaaggct agagccacca tgattgaaca 3061 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg 3121 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg 3181 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc 3241 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt 3301 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc 3361 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca 3421 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc 3481 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg 3541 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct 3601 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc 3661 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc 3721 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta 3781 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt 3841 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga 3901 tggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgaatc 3961 gatagcgata aggatcgggg atgcatcgat aaaataaaag attttattta gtctccagaa 4021 aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagagaacc atcagatgtt 4081 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt 4141 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc 4201 cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tgtatccaat 4261 aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct 4321 gagtgattga ctacccgtca gcgggggtct ttcatgggta acagtttctt gaagttggag 4381 aacaacattc tgagggtagg agtcgaatat taagtaatcc tgactcaatt agccactgtt 4441 ttgaatccac atactccaat actcctgaaa tagttcatta tggacagcgc agaaagagct 4501 ggggagaatt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 4561 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 4621 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 4681 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 4741 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 4801 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 4861 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 4921 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 4981 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 5041 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 5101 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 5161 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 5221 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 5281 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 5341 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 5401 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 5461 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 5521 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 5581 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 5641 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat 5701 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct 5761 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt 5821 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat 5881 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta 5941 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg 6001 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt 6061 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg 6121 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg 6181 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc 6241 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa 6301 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac 6361 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt 6421 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg 6481 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa 6541 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata 6601 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca 6661 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc 6721 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt 6781 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg 6841 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata 6901 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc 6961 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc 7021 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc 7081 agtcacgacg ttgtaaaacg acggcgcaag gaatggtgca tgcaaggaga tggcgcccaa 7141 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc 7201 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc 7261 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggcgat tagtccaatt 7321 tgttaaagac aggatatcag tggtccaggc tctagttttg actcaacaat atcaccagct 7381 gaagcctata gagtacgagc catagataaa ataaaagatt ttatttagtc tccagaaaaa 7441 ggggggaa //