LOCUS Exported 8323 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS p155 pCMV-EGFP/beta-gal SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8323) AUTHORS Kaczmarczyk SJ, Green JE TITLE A single vector containing modified cre recombinase and LOX recombination sequences for inducible tissue-specific amplification of gene expression. JOURNAL Nucleic Acids Res 2001 Jun 15;29(12):E56-6. PUBMED 11410679 REFERENCE 2 (bases 1 to 8323) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8323 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 50..69 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 277..294 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 278..294 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" enhancer 321..624 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 625..828 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 779..799 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 825..849 /label=LNCX /note="Human CMV promoter, forward primer" intron 966..1062 /label=SV40 intron /note="modified SV40 intron with splice donor and acceptor sites" protein_bind 1355..1388 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." regulatory 1397..1406 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1403..2119 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 1403..2119 /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(1448..1469) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1709..1728) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2056..2077 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 2258..2379 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" protein_bind 2386..2419 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS 2512..5559 /codon_start=1 /gene="lacZ" /product="beta-galactosidase" /label=lacZ /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS LNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNPP FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ PNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA ALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP ARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGLR CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD SWSPSVSAEFQLSAGRYHYQLVWCQK" polyA_signal 5823..5957 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6001..6017) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6001..6017) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6014..6036) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6025..6041 /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(6049..6079) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6094..6115 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6232..6249) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6403..6991) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6483..6502) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7162..8022) /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 7785..7804 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8023..8127) /gene="bla" /label=AmpR promoter primer_bind 8195..8213 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(8251..8273) /label=pGEX 3' /note="pGEX vectors, reverse primer" ORIGIN 1 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag 61 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga 121 aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg 181 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta 241 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattc 301 gagcttgcat gcctgcaggt cgttacataa cttacggtaa atggcccgcc tggctgaccg 361 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata 421 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta 481 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc 541 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac 601 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga 661 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg 721 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg 781 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac 841 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac 901 cgatccagcc tccggactct agaggatccg gtactcgagg aactgaaaaa ccagaaagtt 961 aactggtaag tttagtcttt ttgtctttta tttcaggtcc cggatccggt ggtggtgcaa 1021 atcaaagaac tgctcctcag tggatgttgc ctttacttct aggcctgtac ggaagtgtta 1081 cttctgctct aaaagctgcg gaattgtacc cgcggccgca attcccccta ggcttttgca 1141 aaaagcttgg gatctctata atctcgcgca acctattttc ccctcgaaca ctttttaagc 1201 cgtagataaa caggctggga cacttcacat gagcgaaaaa tacatcgtca cctgggacat 1261 gttgcagatc catgcacgta aactcgcaag ccgactgatg ccttctgaac aatggaaagg 1321 cattattgcc gtaagccgtg gcggtctggt accgataact tcgtatagca tacattatac 1381 gaagttatca ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt 1441 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga 1501 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa 1561 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag 1621 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta 1681 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt 1741 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga 1801 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat 1861 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga 1921 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc 1981 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa 2041 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg 2101 catggacgag ctgtacaagt ccggactcag atccaccgga tctagataac tgatcataat 2161 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct 2221 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa 2281 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca 2341 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtataac ttcgtatagc 2401 atacattata cgaagttatc ggtaccggtg ggtgaagacc agaaacagca cctcgaactg 2461 agccgcgata ttgcccagcg tttcaacgcg ctgtatggcg agatcgatcc cgtcgtttta 2521 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc 2581 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg 2641 cgcagcctga atggcgaatg gcgctttgcc tggtttccgg caccagaagc ggtgccggaa 2701 agctggctgg agtgcgatct tcctgaggcc gatactgtcg tcgtcccctc aaactggcag 2761 atgcacggtt acgatgcgcc catctacacc aacgtgacct atcccattac ggtcaatccg 2821 ccgtttgttc ccacggagaa tccgacgggt tgttactcgc tcacatttaa tgttgatgaa 2881 agctggctac aggaaggcca gacgcgaatt atttttgatg gcgttaactc ggcgtttcat 2941 ctgtggtgca acgggcgctg ggtcggttac ggccaggaca gtcgtttgcc gtctgaattt 3001 gacctgagcg catttttacg cgccggagaa aaccgcctcg cggtgatggt gctgcgctgg 3061 agtgacggca gttatctgga agatcaggat atgtggcgga tgagcggcat tttccgtgac 3121 gtctcgttgc tgcataaacc gactacacaa atcagcgatt tccatgttgc cactcgcttt 3181 aatgatgatt tcagccgcgc tgtactggag gctgaagttc agatgtgcgg cgagttgcgt 3241 gactacctac gggtaacagt ttctttatgg cagggtgaaa cgcaggtcgc cagcggcacc 3301 gcgcctttcg gcggtgaaat tatcgatgag cgtggtggtt atgccgatcg cgtcacacta 3361 cgtctgaacg tcgaaaaccc gaaactgtgg agcgccgaaa tcccgaatct ctatcgtgcg 3421 gtggttgaac tgcacaccgc cgacggcacg ctgattgaag cagaagcctg cgatgtcggt 3481 ttccgcgagg tgcggattga aaatggtctg ctgctgctga acggcaagcc gttgctgatt 3541 cgaggcgtta accgtcacga gcatcatcct ctgcatggtc aggtcatgga tgagcagacg 3601 atggtgcagg atatcctgct gatgaagcag aacaacttta acgccgtgcg ctgttcgcat 3661 tatccgaacc atccgctgtg gtacacgctg tgcgaccgct acggcctgta tgtggtggat 3721 gaagccaata ttgaaaccca cggcatggtg ccaatgaatc gtctgaccga tgatccgcgc 3781 tggctaccgg cgatgagcga acgcgtaacg cgaatggtgc agcgcgatcg taatcacccg 3841 agtgtgatca tctggtcgct ggggaatgaa tcaggccacg gcgctaatca cgacgcgctg 3901 tatcgctgga tcaaatctgt cgatccttcc cgcccggtgc agtatgaagg cggcggagcc 3961 gacaccacgg ccaccgatat tatttgcccg atgtacgcgc gcgtggatga agaccagccc 4021 ttcccggctg tgccgaaatg gtccatcaaa aaatggcttt cgctacctgg agagacgcgc 4081 ccgctgatcc tttgcgaata cgcccacgcg atgggtaaca gtcttggcgg tttcgctaaa 4141 tactggcagg cgtttcgtca gtatccccgt ttacagggcg gcttcgtctg ggactgggtg 4201 gatcagtcgc tgattaaata tgatgaaaac ggcaacccgt ggtcggctta cggcggtgat 4261 tttggcgata cgccgaacga tcgccagttc tgtatgaacg gtctggtctt tgccgaccgc 4321 acgccgcatc cagcgctgac ggaagcaaaa caccagcagc agtttttcca gttccgttta 4381 tccgggcaaa ccatcgaagt gaccagcgaa tacctgttcc gtcatagcga taacgagctc 4441 ctgcactgga tggtggcgct ggatggtaag ccgctggcaa gcggtgaagt gcctctggat 4501 gtcgctccac aaggtaaaca gttgattgaa ctgcctgaac taccgcagcc ggagagcgcc 4561 gggcaactct ggctcacagt acgcgtagtg caaccgaacg cgaccgcatg gtcagaagcc 4621 gggcacatca gcgcctggca gcagtggcgt ctggcggaaa acctcagtgt gacgctcccc 4681 gccgcgtccc acgccatccc gcatctgacc accagcgaaa tggatttttg catcgagctg 4741 ggtaataagc gttggcaatt taaccgccag tcaggctttc tttcacagat gtggattggc 4801 gataaaaaac aactgctgac gccgctgcgc gatcagttca cccgtgcacc gctggataac 4861 gacattggcg taagtgaagc gacccgcatt gaccctaacg cctgggtcga acgctggaag 4921 gcggcgggcc attaccaggc cgaagcagcg ttgttgcagt gcacggcaga tacacttgct 4981 gatgcggtgc tgattacgac cgctcacgcg tggcagcatc aggggaaaac cttatttatc 5041 agccggaaaa cctaccggat tgatggtagt ggtcaaatgg cgattaccgt tgatgttgaa 5101 gtggcgagcg atacaccgca tccggcgcgg attggcctga actgccagct ggcgcaggta 5161 gcagagcggg taaactggct cggattaggg ccgcaagaaa actatcccga ccgccttact 5221 gccgcctgtt ttgaccgctg ggatctgcca ttgtcagaca tgtatacccc gtacgtcttc 5281 ccgagcgaaa acggtctgcg ctgcgggacg cgcgaattga attatggccc acaccagtgg 5341 cgcggcgact tccagttcaa catcagccgc tacagtcaac agcaactgat ggaaaccagc 5401 catcgccatc tgctgcacgc ggaagaaggc acatggctga atatcgacgg tttccatatg 5461 gggattggtg gcgacgactc ctggagcccg tcagtatcgg cggaattcca gctgagcgcc 5521 ggtcgctacc attaccagtt ggtctggtgt caaaaataat aataaccggg caggccatgt 5581 ctgcccgtat ttcgcgtaag gaaatccatt atgtactatt taaaaaacac aaacttttgg 5641 atgttcggtt tattcttttt cttttacttt tttatcatgg gagcctactt cccgtttttc 5701 ccgatttggc tacatgacat caaccatatc agcaaaagtg atacgggtat tatttttgcc 5761 gctatttctc tgttctcgct attattccaa ccgctgtttg gtctgctttc tgacaaactc 5821 ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca 5881 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat 5941 cttatcatgt ctggatcctc tagagtcgac ctgcaggcat gcaagcttgg cgtaatcatg 6001 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 6061 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 6121 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 6181 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 6241 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 6301 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 6361 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 6421 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 6481 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 6541 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 6601 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 6661 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 6721 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 6781 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 6841 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 6901 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 6961 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 7021 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 7081 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 7141 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 7201 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 7261 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 7321 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 7381 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 7441 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 7501 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 7561 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 7621 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 7681 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 7741 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 7801 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 7861 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 7921 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 7981 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 8041 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 8101 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta 8161 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg 8221 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt 8281 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgt //