LOCUS Exported 7369 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pNrl-Cre SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7369) AUTHORS Matsuda T, Cepko CL TITLE Controlled expression of transgenes introduced by in vivo electroporation. JOURNAL Proc Natl Acad Sci U S A. 2007 Jan 5. ():. PUBMED 17209010 REFERENCE 2 (bases 1 to 7369) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7369 /organism="synthetic DNA construct" /mol_type="other DNA" regulatory 3229..3238 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3235..4263 /codon_start=1 /product="site-specific recombinase" /label=Cre /note="Cre is a site-specific recombinase from bacteriophage P1. Recombination occurs at loxP sequences." /translation="MANLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE DGD" CDS 4273..4302 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" primer_bind complement(4393..4412) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 4458..4513 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(4512..4531) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(4872..4888) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4872..4888) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4885..4907) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4896..4912 /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(4920..4950) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4965..4986 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5045..5240 /label=SV40 promoter /note="SV40 early promoter" rep_origin 5091..5226 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5153..5172 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 5246..5380 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5283..5302) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5337..5356 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5448..5465) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5619..6207) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5699..5718) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6378..7238) /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 7001..7020 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7239..7343) /gene="bla" /label=AmpR promoter ORIGIN 1 gtcgacttac atgtacatac ttcataatgg tctagaaggg cttattcaaa tttggcgttt 61 actcagagtc ctcggggctc ttatccggga tgtggtgatg atgtgaggag aggcaactct 121 caaactagga ccatcactcg gggcttgtgt atcagtagtg aggtccagca atggaaggga 181 aatcccggga aaaaagtcta tgtgatccct aggagcaggt ctgagctctt tgtgcagaaa 241 atttctggat acctggagta tgattctcag ggaccagggg tctgggtcag agaagggtgg 301 ctgtctcaca tcatggggta cattcagcta tagaaagacc agcatgaacc aggcagtggt 361 acctggcctg aaccaggact ggctcagctt gcttgagctt atttggcccg actcccttca 421 ctccttagca gcttagctct cccacctctc tcctgagtcc tccagtaaac aagaggatca 481 ggacagatat gacagctgcc tggtggggca agtgtctggt caggtcaaca gcaggcatga 541 agggggctac tacactaaat gctgccaacc tttttccttt agggaagttc taggatctct 601 ttcccggata tccactgtaa aacctcattc cactggcatg tatccctatt cctttcagca 661 tccctattgc ccttttgtgg atggttcctg gccttacatt ttccgcttct tttaagaaaa 721 taagcaacag gtcttggtga agagtaacat ttccattcga atagtgactc tatttcataa 781 accaaacgaa acgaaacgaa acgaaacgaa acgaaacaaa acaaaacaaa acaaaacacc 841 aagagacctg ggcatgggag agttgtgata tctgacctaa gccttctaat gtatgtagag 901 gaagcttgga ttccaaatgt cctccaaagc tgggcatggt ggtgcgtgcc tttacgccct 961 gtactaggaa ggcacaagca ggtagatcta tgggttcaag gtcaatctgg tctacaaaga 1021 gagttctagg acggctaggg ctacacagag aaaccctgtt ttgaaaacca aaccaaacca 1081 aaccaatgtc ctccaaaggc atttgagttc aaaacttgtc attattggaa ggtggtgaaa 1141 ttttcaagag gtggggccct gtgggaagtc ttctggtctt tgggttttcc ctgaaggaag 1201 caaaggccta acccctccct ggtcatcggt tggtaccctt gagggaagca ggagtcctat 1261 ccacttccct tcccttttct tttttttttt ttttgtccca gtgataaggt gagtagcttt 1321 cagttgtgtc ctcgccataa cacttgactc actacaggca atacatgatg ggaccaactg 1381 ctcactggct aggctctcca aggcagcaaa ccaaagcaaa gcgctttccg attaagtcaa 1441 ttacttcagg tatgtattgg agcaattatg ttagtgttag tcggtttttg ttattgtgac 1501 aaagtgcccg agaaagatca aaggagggga gccttctttg ggctaatggc ttcagtttca 1561 gctgatggtt gtgggtgtcc tattgctttg gtccaaaatc taacatcacg atggaaaatg 1621 tatagtgaag aagtaaagag agaaaagaga gaggaagagg gggaagtgga ggtgggggag 1681 aaggagggat ggagggacac agacaaaatg caccaatgaa ctaaggagaa gatacaccat 1741 tcaaacgcat gttttttcta gttaggccaa ttcatactga cagccaattc agtatggatt 1801 cttcagctgg ttagttatgg ataagtttaa cctctaataa tatctcagtg aaaactagct 1861 tttaaaatat accggaacca tagcagtgat agaaaactaa tgtagggtgg acacacacac 1921 acacacacac acacaaacac acactttgag actgggttga aggtaggcat ctgtggacca 1981 gtgggtctca aacattagtg tgtttcagag ctgttggaag ggtttattaa acagcagctg 2041 tctataacct acttactgag tttctggttc tgtgagtttg aggtaaagca aaagaatttg 2101 tttcttcaag ttcccaaggg gtgatttctt aagactgtgg aagccattgt tccaacaggc 2161 agggaatacc ctttaacttc tggagtttct tttatcacag cacctattta aacagcttca 2221 ggcaggttta aacatctcct tggctaagtc ctactatccc tgcttcggcc cctactaggg 2281 ggtgacagct ctaagaggcg ttaggcgatt aggctgaaaa tgtaggtcac accccagccg 2341 tctgggaatg agcgagagag gagaagaggg actgagacac cgcacgggga aaaggtgaga 2401 ggaagctgag ggctcatgag ggcattcaca gaagtgtggt ggtgattgga tttcagcttc 2461 ttttagtctc cagatgaaag accagagttg ggatctccgc tttttctatc tcttctctcc 2521 agctgggctg cttgtcttcg ggatgggcta ttaagagcga gtttcttcat ggaagctcat 2581 ggcttatctt tctaatacag tgactctgtg aggaataaga cgagtcaggg tgaaggtagg 2641 gcaggggctg aaatgtgagg atgcttgaaa agaagcagca gccccaacaa gctggctgag 2701 gatgggctgg gaacgcgtat atgtctcctt gacacggagt cttaaaagag gttttgggta 2761 gtatgtctgg actcttacca ctctaagctt tgagtgattg ggctgagaat tgtgtttcca 2821 tggaggggtg gtgtttgggg tcaacatggc tctctatatt ggaaaccaag ctcacaacat 2881 aagcaaggtc tgatatagga agcttctctg gggaagatgc cgtttgtttt tttccttttt 2941 ggagaactga tagcacctgt ctttcactgg cttctgagtc cctgccccag gccaatgaga 3001 tatgcaaata agcccttcat cttattggat gaggctcgag gaccacagat gatctcaggg 3061 aaccagtcct ttaagagtgt cctctgggct ctgtgcccac agcccccttg ttctgaatac 3121 agggacgaca ccagcccctg ctctatggag tatttagtct ccagggaagc tgtgcctttc 3181 tggttctgac agtgactacg tcatctctgc cattacgaat tctgagccgc caccatggcc 3241 aatttactga ccgtacacca aaatttgcct gcattaccgg tcgatgcaac gagtgatgag 3301 gttcgcaaga acctgatgga catgttcagg gatcgccagg cgttttctga gcatacctgg 3361 aaaatgcttc tgtccgtttg ccggtcgtgg gcggcatggt gcaagttgaa taaccggaaa 3421 tggtttcccg cagaacctga agatgttcgc gattatcttc tatatcttca ggcgcgcggt 3481 ctggcagtaa aaactatcca gcaacatttg ggccagctaa acatgcttca tcgtcggtcc 3541 gggctgccac gaccaagtga cagcaatgct gtttcactgg ttatgcggcg gatccgaaaa 3601 gaaaacgttg atgccggtga acgtgcaaaa caggctctag cgttcgaacg cactgatttc 3661 gaccaggttc gttcactcat ggaaaatagc gatcgctgcc aggatatacg taatctggca 3721 tttctgggga ttgcttataa caccctgtta cgtatagccg aaattgccag gatcagggtt 3781 aaagatatct cacgtactga cggtgggaga atgttaatcc atattggcag aacgaaaacg 3841 ctggttagca ccgcaggtgt agagaaggca cttagcctgg gggtaactaa actggtcgag 3901 cgatggattt ccgtctctgg tgtagctgat gatccgaata actacctgtt ttgccgggtc 3961 agaaaaaatg gtgttgccgc gccatctgcc accagccagc tatcaactcg cgccctggaa 4021 gggatttttg aagcaactca tcgattgatt tacggcgcta aggatgactc tggtcagaga 4081 tacctggcct ggtctggaca cagtgcccgt gtcggagccg cgcgagatat ggcccgcgct 4141 ggagtttcaa taccggagat catgcaagct ggtggctgga ccaatgtaaa tattgtcatg 4201 aactatatcc gtaacctgga tagtgaaaca ggggcaatgg tgcgcctgct ggaagatggc 4261 gatggaccgg tggaacaaaa acttatttct gaagaagatc tgtgatagcg gccgcactcc 4321 tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa 4381 ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga 4441 gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt 4501 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga 4561 gtatttggtt tagagtttgg caacatatgc catatgctgg ctgccatgaa caaaggtggc 4621 tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa 4681 aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa 4741 catccctaaa attttcctta catgttttac tagccagatt tttcctcctc tcctgactac 4801 tcccagtcat agctgtccct cttctcttat gaagatccct cgacctgcag cccaagcttg 4861 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 4921 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 4981 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg 5041 atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc 5101 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg 5161 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg 5221 gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg gttacaaata 5281 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg 5341 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta atgaatcggc 5401 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac 5461 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata 5521 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa 5581 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 5641 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 5701 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 5761 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca 5821 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 5881 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 5941 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 6001 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg 6061 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 6121 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 6181 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 6241 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc 6301 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag 6361 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt 6421 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag 6481 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca 6541 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact 6601 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca 6661 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg 6721 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc 6781 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg 6841 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca 6901 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt 6961 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc 7021 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc 7081 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca 7141 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa 7201 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat 7261 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa 7321 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctg //