LOCUS Exported 8656 bp ds-DNA circular SYN 06-MAR-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8656) AUTHORS MReijns TITLE Direct Submission JOURNAL Exported Mar 6, 2018 from SnapGene Viewer 4.1.7 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8656 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 580..921 /label=MESV Psi /note="packaging signal of murine embryonic stem cell virus" CDS 988..1404 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" CDS 1429..2145 /codon_start=1 /product="enhanced GFP" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" protein_bind 2218..2242 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2245..3813 /codon_start=1 /label=cGAS /translation="MQPWHGKAMQRASEAGATAPKASARNARGAPMDPTESPAAPEAAL PKAGKFGPARKSGSRQKKSAPDTQERPPVRATGARAKKAPQRAQDTQPSDATSAPGAEG LEPPAAREPALSRAGSCRQRGARCSTKPRPPPGPWDVPSPGLPVSAPILVRRDAAPGAS KLRAVLEKLKLSRDDISTAAGMVKGVVDHLLLRLKCDSAFRGVGLLNTGSYYEHVKISA PNEFDVMFKLEVPRIQLEEYSNTRAYYFVKFKRNPKENPLSQFLEGEILSASKMLSKFR KIIKEEINDIKDTDVIMKRKRGGSPAVTLLISEKISVDITLALESKSSWPASTQEGLRI QNWLSAKVRKQLRLKPFYLVPKHAKEGNGFQEETWRLSFSHIEKEILNNHGKSKTCCEN KEEKCCRKDCLKLMKYLLEQLKERFKDKKHLDKFSSYHVKTAFFHVCTQNPQDSQWDRK DLGLCFDNCVTYFLQCLRTEKLENYFIPEFNLFSSNLIDKRSKEFLTKQIEYERNNEFP VFDEF" promoter 3830..4329 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" CDS 4350..4949 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" LTR 5049..5563 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus" protein_bind 5725..5741 /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(5749..5779) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5794..5815 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(6103..6691) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6862..7722) /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" promoter complement(7723..7827) /gene="bla" /label=AmpR promoter ORIGIN 1 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca 61 tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc 121 agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag 181 aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt 241 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt 301 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc 361 cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tattcccaat 421 aaagcctctt gctgtttgca tccgaatcgt ggactcgctg atccttggga gggtctcctc 481 agattgattg actgcccacc tcgggggtct ttcatttgga ggttccaccg agatttggag 541 acccctgccc agggaccacc gacccccccg ccgggaggta agctggccag cggtcgtttc 601 gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg 661 tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttctga 721 acacccggcc gcaaccctgg gagacgtccc agggactttg ggggccgttt ttgtggcccg 781 acctgaggaa gggagtcgat gtggaatccg accccgtcag gatatgtggt tctggtagga 841 gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttggaaccga 901 agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact 961 gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc 1021 ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag 1081 agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga 1141 gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc 1201 ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac 1261 ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc 1321 gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca 1381 gccctcactc cttctctagg cgccggaatt agatctctcg aggttaacat ggtgagcaag 1441 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac 1501 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc 1561 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc 1621 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc 1681 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac 1741 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc 1801 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac 1861 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg 1921 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag 1981 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc 2041 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc 2101 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtccgg ccggactcag 2161 atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg gatcatcaca 2221 agtttgtaca aaaaagcagg cttcatgcag ccttggcacg gaaaggccat gcagagagct 2281 tccgaggccg gagccactgc ccccaaggct tccgcacgga atgccagggg cgccccgatg 2341 gatcccaccg agtctccggc tgcccccgag gccgccctgc ctaaggcggg aaagttcggc 2401 cccgccagga agtcgggatc ccggcagaaa aagagcgccc cggacaccca ggagaggccg 2461 cccgtccgcg caactggggc ccgcgccaaa aaggcccctc agcgcgccca ggacacgcag 2521 ccgtctgacg ccaccagcgc ccctggggca gaggggctgg agcctcctgc ggctcgggag 2581 ccggctcttt ccagggctgg ttcttgccgc cagaggggcg cgcgctgctc cacgaagcca 2641 agacctccgc ccgggccctg ggacgtgccc agccccggcc tgccggtctc ggcccccatt 2701 ctcgtacgga gggatgcggc gcctggggcc tcgaagctcc gggcggtttt ggagaagttg 2761 aagctcagcc gcgatgatat ctccacggcg gcggggatgg tgaaaggggt tgtggaccac 2821 ctgctgctca gactgaagtg cgactccgcg ttcagaggcg tcgggctgct gaacaccggg 2881 agctactatg agcacgtgaa gatttctgca cctaatgaat ttgatgtcat gtttaaactg 2941 gaagtcccca gaattcaact agaagaatat tccaacactc gtgcatatta ctttgtgaaa 3001 tttaaaagaa atccgaaaga aaatcctctg agtcagtttt tagaaggtga aatattatca 3061 gcttctaaga tgctgtcaaa gtttaggaaa atcattaagg aagaaattaa cgacattaaa 3121 gatacagatg tcatcatgaa gaggaaaaga ggagggagcc ctgctgtaac acttcttatt 3181 agtgaaaaaa tatctgtgga tataaccctg gctttggaat caaaaagtag ctggcctgct 3241 agcacccaag aaggcctgcg cattcaaaac tggctttcag caaaagttag gaagcaacta 3301 cgactaaagc cattttacct tgtacccaag catgcaaagg aaggaaatgg tttccaagaa 3361 gaaacatggc ggctatcctt ctctcacatc gaaaaggaaa ttttgaacaa tcatggaaaa 3421 tctaaaacgt gctgtgaaaa caaagaagag aaatgttgca ggaaagattg tttaaaacta 3481 atgaaatacc ttttagaaca gctgaaagaa aggtttaaag acaaaaaaca tctggataaa 3541 ttctcttctt atcatgtgaa aactgccttc tttcacgtat gtacccagaa ccctcaagac 3601 agtcagtggg accgcaaaga cctgggcctc tgctttgata actgcgtgac atactttctt 3661 cagtgcctca ggacagaaaa acttgagaat tattttattc ctgaattcaa tctattctct 3721 agcaacttaa ttgacaaaag aagtaargaa tttctgacaa agcaaattga atatgaaaga 3781 aacaatgagt ttccagtttt tgatgaattt tgagttaacg aattctaccg ggtaggggag 3841 gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc 3901 tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct 3961 ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc 4021 cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt 4081 ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc 4141 caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg 4201 ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg 4261 gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg 4321 ggcctttcga cctgcagccc aagcttacca tgaccgagta caagcccacg gtgcgcctcg 4381 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc 4441 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag 4501 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg 4561 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga 4621 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag 4681 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct 4741 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg 4801 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct 4861 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct 4921 ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg 4981 aaaggagcgc acgaccccat gcatcgataa aataaaagat tttatttagt ctccagaaaa 5041 aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt 5101 gcaaggcatg gaaaatacat aactgagaat agagaagttc agatcaaggt taggaacaga 5161 gagacagcag aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca 5221 gggccaagaa cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat 5281 cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc 5341 aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc 5401 ccacaacccc tcactcggcg cgccagtcct ccgatagact gcgtcgcccg ggtacccgtg 5461 tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg 5521 tctcctctga gtgattgact acccgtcagc gggggtcttt catgggtaac agtttcttga 5581 agttggagaa caacattctg agggtaggag tcgaatatta agtaatcctg actcaattag 5641 ccactgtttt gaatccacat actccaatac tcctgaaata gttcattatg gacagcgcag 5701 aaagagctgg ggagaattgt gaaattgtta tccgctcaca attccacaca acatacgagc 5761 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 5821 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 5881 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 5941 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 6001 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 6061 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 6121 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 6181 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 6241 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 6301 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 6361 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 6421 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 6481 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 6541 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 6601 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 6661 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 6721 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 6781 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 6841 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 6901 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 6961 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 7021 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 7081 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 7141 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 7201 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 7261 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 7321 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 7381 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 7441 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 7501 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 7561 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 7621 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 7681 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 7741 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 7801 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta 7861 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg 7921 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt 7981 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg 8041 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt 8101 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg 8161 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 8221 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 8281 gttttcccag tcacgacgtt gtaaaacgac ggcgcaagga atggtgcatg caaggagatg 8341 gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc 8401 atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca 8461 gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggcgatta 8521 gtccaatttg ttaaagacag gatatcagtg gtccaggctc tagttttgac tcaacaatat 8581 caccagctga agcctataga gtacgagcca tagataaaat aaaagatttt atttagtctc 8641 cagaaaaagg ggggaa //