LOCUS Exported 6144 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION pMVP destination vector, empty PiggyBac transposon vector backbone w/ eGFP selection cassette. ACCESSION . VERSION . KEYWORDS LB601: pMVP/PB/eGFP-DEST SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6144) AUTHORS Haldeman JM, Conway AE, Arlotto ME, Slentz DH, Muoio DM, Becker TC, Newgard CB TITLE Creation of versatile cloning platforms for transgene expression and dCas9-based epigenome editing. JOURNAL Nucleic Acids Res. 2018 Dec 27. pii: 5264291. doi: 10.1093/nar/gky1286. PUBMED 30590691 REFERENCE 2 (bases 1 to 6144) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6144 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 24..128 /gene="bla" /label=AmpR promoter CDS 129..989 /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 complement(347..366) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1160..1748 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1649..1668 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 1921..1938 /label=L4440 /note="L4440 vector, forward primer" primer_bind 1944..1961 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 1945..1961 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" repeat_region 1987..2049 /label=piggyBac right (3') inverted repeat /note="piggyBac transposon-specific inverted terminal repeat sequence (ITR)" protein_bind 2263..2387 /gene="mutant version of attR" /label=attR1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" promoter 2412..2442 /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" CDS 2496..3155 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(2562..2581) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" CDS 3497..3802 /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /label=ccdB /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" primer_bind 3681..3700 /label=ccdB-fwd /note="ccdB gene, forward primer" protein_bind complement(3843..3967) /gene="mutant version of attR" /label=attR2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(3982..3998) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3983..3999) /label=pBluescriptKS /note="For pBluescript vector" promoter 4019..4335 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4186..4321 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4248..4267 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 4383..4430 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 4449..5168 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 4449..5165 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(4494..4515) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4755..4774) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 5102..5123 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(5208..5227) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5252..5300 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" primer_bind complement(5516..5537) /label=F1ori-F /note="F1 origin, forward primer" repeat_region 5821..5855 /label=piggyBac left (5') inverted repeat /note="piggyBac transposon-specific inverted terminal repeat sequence (ITR)" primer_bind 6035..6054 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt 61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa 121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt 181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt 241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt 301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg 361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga 421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa 481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga 541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa 601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca 661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta 721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac 781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc 841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag 901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga 961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt 1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata 1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag 1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa 1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt 1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc 1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa 1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa 1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc 1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa 1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa 1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg 1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc 1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg 1801 ctcacatgct gtacagctta agtcatgttc tttcctgcgt tatcccctga ttctgtggat 1861 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc 1921 agcgagtcag tgagcgagga agctgtaaaa cgacggccag tagctccacc gccgcagcta 1981 gattaaccct agaaagataa tcatattgtg acgtacgtta aagataatca tgcgtaaaat 2041 tgacgcatgt gttttatcgg tctgtatatc gaggtttatt tattaatttg aatagatatt 2101 aagttttatt atatttacac ttacatacta ataataaatt caacaaacaa tttatttatg 2161 tttatttatt tattaaaaaa aaacaaaaac tcaaaatttc ttctataaag taacaaaact 2221 tttatgacta gtggatcccc cgggctgcag gaattcgata tcacaagttt gtacaaaaaa 2281 gctgaacgag aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa 2341 aaacagacta cataatactg taaaacacaa catatccagt cactatggcg gccgcattag 2401 gcaccccagg ctttacactt tatgcttccg gctcgtataa tgtgtggatt ttgagttagg 2461 atccgtcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata 2521 ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg 2581 ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa 2641 agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg 2701 ctcatccgga attccgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc 2761 acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat 2821 accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg 2881 aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc 2941 cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc 3001 ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga 3061 ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt aatgaattac 3121 aacagtactg cgatgagtgg cagggcgggg cgtaaacgcg tggatccggc ttactaaaag 3181 ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg tataagaata tatactgata 3241 tgtatacccg aagtatgtca aaaagaggta tgctatgaag cagcgtatta cagtgacagt 3301 tgacagcgac agctatcagt tgctcaaggc atatatgatg tcaatatctc cggtctggta 3361 agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat 3421 caggaaggga tggctgaggt cgcccggttt attgaaatga acggctcttt tgctgacgag 3481 aacaggggct ggtgaaatgc agtttaaggt ttacacctat aaaagagaga gccgttatcg 3541 tctgtttgtg gatgtacaga gtgatattat tgacacgccc gggcgacgga tggtgatccc 3601 cctggccagt gcacgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca 3661 tatcggggat gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtctccgt 3721 tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa 3781 cctgatgttc tggggaatat aaatgtcagg ctcccttata cacagccagt ctgcaggtcg 3841 accatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa 3901 tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcagct ttcttgtaca 3961 aagtggtgat atcaagctta tcgataccgt cgacctcgac atggtcatag ctgttgacgg 4021 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 4081 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 4141 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact 4201 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 4261 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 4321 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac 4381 gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga 4441 actaaaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg 4501 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg 4561 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct 4621 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc 4681 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca 4741 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg 4801 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc 4861 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc 4921 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc 4981 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg 5041 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc 5101 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt 5161 acaagtaacg taccggttag taatgagttt aaacggggga ggctaactga aacacggaag 5221 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac 5281 gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga 5341 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca 5401 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc 5461 catagcagat ccggttaaat caaaagaata gaccgagata gggttgagtg ttgttccagt 5521 ttggaacaag agtccacgct gtccctgata tctataacaa gaaaatatat atataataag 5581 ttatcacgta agtagaacat gaaataacaa tataattatc gtatgagtta aatcttaaaa 5641 gtcacgtaaa agataatcat gcgtcatttt gactcacgcg gtcgttatag ttcaaaatca 5701 gtgacactta ccgcattgac aagcacgcct cacgggagct ccaagcggcg actgagatgt 5761 cctaaatgca cagcgacgga ttcgcgctat ttagaaagag agagcaatat ttcaagaatg 5821 catgcgtcaa ttttacgcag actatctttc tagggttaat ctagctgcat caagggcgaa 5881 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg 5941 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt 6001 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg 6061 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta 6121 atgcgccgct acagggcgcg tcag //