LOCUS Exported 7350 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS MSCV IRES Luciferase SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7350) TITLE Lowe Lab Plasmids REFERENCE 2 (bases 1 to 7350) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7350 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(23..45) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 145..164 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 358..380 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 508..527 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" LTR 727..1243 /label=5' LTR /note="5' long terminal repeat from murine embryonic stem cell virus" misc_feature 1307..1648 /label=MESV Psi /note="packaging signal of murine embryonic stem cell virus" CDS 1715..2131 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 2061..2083 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 2099..2115 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" misc_feature 2177..2729 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(2321..2338) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 2548..2567 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 2741..4393 /codon_start=1 /gene="luc+" /product="firefly luciferase" /label=luciferase /note="enhanced luc+ version of the luciferase gene" /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV" primer_bind complement(2807..2825) /label=LucNrev /note="Firefly luciferase, reverse primer" LTR 4471..4985 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus" protein_bind 5147..5163 /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(5171..5201) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5216..5237 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5354..5371) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5525..6113) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5605..5624) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6284..7144) /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 6907..6926 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7145..7249) /gene="bla" /label=AmpR promoter primer_bind 7317..7335 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt 61 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg 121 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata 181 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc 241 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc 301 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc 361 agtcacgacg ttgtaaaacg acggcgcaag gaatggtgca tgcaaggaga tggcgcccaa 421 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc 481 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc 541 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggcgat tagtccaatt 601 tgttaaagac aggatatcag tggtccaggc tctagttttg actcaacaat atcaccagct 661 gaagcctata gagtacgagc catagataaa ataaaagatt ttatttagtc tccagaaaaa 721 ggggggaatg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg 781 caaggcatgg aaaatacata actgagaata gagaagttca gatcaaggtt aggaacagag 841 agacagcaga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag 901 ggccaagaac agatggtccc cagatgcggt cccgccctca gcagtttcta gagaaccatc 961 agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca 1021 atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc 1081 cacaacccct cactcggcgc gccagtcctc cgatagactg cgtcgcccgg gtacccgtat 1141 tcccaataaa gcctcttgct gtttgcatcc gaatcgtgga ctcgctgatc cttgggaggg 1201 tctcctcaga ttgattgact gcccacctcg ggggtctttc atttggaggt tccaccgaga 1261 tttggagacc cctgcccagg gaccaccgac ccccccgccg ggaggtaagc tggccagcgg 1321 tcgtttcgtg tctgtctctg tctttgtgcg tgtttgtgcc ggcatctaat gtttgcgcct 1381 gcgtctgtac tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga 1441 gttctgaaca cccggccgca accctgggag acgtcccagg gactttgggg gccgtttttg 1501 tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat atgtggttct 1561 ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg 1621 gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt gttgtctctg 1681 tctgactgtg tttctgtatt tgtctgaaaa ttagggccag actgttacca ctcccttaag 1741 tttgacctta ggtcactgga aagatgtcga gcggatcgct cacaaccagt cggtagatgt 1801 caagaagaga cgttgggtta ccttctgctc tgcagaatgg ccaaccttta acgtcggatg 1861 gccgcgagac ggcaccttta accgagacct catcacccag gttaagatca aggtcttttc 1921 acctggcccg catggacacc cagaccaggt cccctacatc gtgacctggg aagccttggc 1981 ttttgacccc cctccctggg tcaagccctt tgtacaccct aagcctccgc ctcctcttcc 2041 tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct 2101 ttatccagcc ctcactcctt ctctaggcgc cggaattaga tctctcgagg ttaacgaatt 2161 ccgccccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt 2221 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct 2281 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa 2341 ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg 2401 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc 2461 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg 2521 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg 2581 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc 2641 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg 2701 gttttccttt gaaaaacacg atgataatat ggccacaacc atggaagacg ccaaaaacat 2761 aaagaaaggc ccggcgccat tctatccgct ggaagatgga accgctggag agcaactgca 2821 taaggctatg aagagatacg ccctggttcc tggaacaatt gcttttacag atgcacatat 2881 cgaggtggac atcacttacg ctgagtactt cgaaatgtcc gttcggttgg cagaagctat 2941 gaaacgatat gggctgaata caaatcacag aatcgtcgta tgcagtgaaa actctcttca 3001 attctttatg ccggtgttgg gcgcgttatt tatcggagtt gcagttgcgc ccgcgaacga 3061 catttataat gaacgtgaat tgctcaacag tatgggcatt tcgcagccta ccgtggtgtt 3121 cgtttccaaa aaggggttgc aaaaaatttt gaacgtgcaa aaaaagctcc caatcatcca 3181 aaaaattatt atcatggatt ctaaaacgga ttaccaggga tttcagtcga tgtacacgtt 3241 cgtcacatct catctacctc ccggttttaa tgaatacgat tttgtgccag agtccttcga 3301 tagggacaag acaattgcac tgatcatgaa ctcctctgga tctactggtc tgcctaaagg 3361 tgtcgctctg cctcatagaa ctgcctgcgt gagattctcg catgccagag atcctatttt 3421 tggcaatcaa atcattccgg atactgcgat tttaagtgtt gttccattcc atcacggttt 3481 tggaatgttt actacactcg gatatttgat atgtggattt cgagtcgtct taatgtatag 3541 atttgaagaa gagctgtttc tgaggagcct tcaggattac aagattcaaa gtgcgctgct 3601 ggtgccaacc ctattctcct tcttcgccaa aagcactctg attgacaaat acgatttatc 3661 taatttacac gaaattgctt ctggtggcgc tcccctctct aaggaagtcg gggaagcggt 3721 tgccaagagg ttccatctgc caggtatcag gcaaggatat gggctcactg agactacatc 3781 agctattctg attacacccg agggggatga taaaccgggc gcggtcggta aagttgttcc 3841 attttttgaa gcgaaggttg tggatctgga taccgggaaa acgctgggcg ttaatcaaag 3901 aggcgaactg tgtgtgagag gtcctatgat tatgtccggt tatgtaaaca atccggaagc 3961 gaccaacgcc ttgattgaca aggatggatg gctacattct ggagacatag cttactggga 4021 cgaagacgaa cacttcttca tcgttgaccg cctgaagtct ctgattaagt acaaaggcta 4081 tcaggtggct cccgctgaat tggaatccat cttgctccaa caccccaaca tcttcgacgc 4141 aggtgtcgca ggtcttcccg acgatgacgc cggtgaactt cccgccgccg ttgttgtttt 4201 ggagcacgga aagacgatga cggaaaaaga gatcgtggat tacgtcgcca gtcaagtaac 4261 aaccgcgaaa aagttgcgcg gaggagttgt gtttgtggac gaagtaccga aaggtcttac 4321 cggaaaactc gacgcaagaa aaatcagaga gatcctcata aaggccaaga agggcggaaa 4381 gatcgccgtg taattctagg atccgtcgac ctgcagccaa gcttatcgat aaaataaaag 4441 attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag 4501 ctagcttaag taacgccatt ttgcaaggca tggaaaatac ataactgaga atagagaagt 4561 tcagatcaag gttaggaaca gagagacagc agaatatggg ccaaacagga tatctgtggt 4621 aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc ggtcccgccc 4681 tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc 4741 ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc 4801 tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc ctccgataga 4861 ctgcgtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg 4921 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct 4981 ttcatgggta acagtttctt gaagttggag aacaacattc tgagggtagg agtcgaatat 5041 taagtaatcc tgactcaatt agccactgtt ttgaatccac atactccaat actcctgaaa 5101 tagttcatta tggacagcgc agaaagagct ggggagaatt gtgaaattgt tatccgctca 5161 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 5221 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 5281 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 5341 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 5401 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 5461 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 5521 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 5581 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 5641 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 5701 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 5761 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 5821 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 5881 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 5941 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag 6001 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 6061 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 6121 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 6181 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 6241 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 6301 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 6361 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 6421 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 6481 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 6541 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 6601 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 6661 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 6721 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 6781 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 6841 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 6901 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 6961 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 7021 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 7081 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 7141 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 7201 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 7261 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata 7321 ggcgtatcac gaggcccttt cgtctcgcgc //