LOCUS Exported 6757 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pTriEx-HTNC SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6757) AUTHORS Peitz M, Pfannkuche K, Rajewsky K, Edenhofer F TITLE Ability of the hydrophobic FGF and basic TAT peptides to promote cellular uptake of recombinant Cre recombinase: a tool for efficient genetic engineering of mammalian genomes. JOURNAL Proc Natl Acad Sci U S A. 2002 Apr 2. 99(7):4489-94. PUBMED 11904364 REFERENCE 2 (bases 1 to 6757) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6757 /organism="synthetic DNA construct" /mol_type="other DNA" misc_recomb 4..831 /label=baculovirus recombination region (lef2/ORF603) /note="contains ORF603 and part of lef2" CDS 4..128 /codon_start=3 /product="baculovirus late expression factor 2" /label=lef2 /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY" CDS complement(166..771) /codon_start=1 /product="baculovirus ORF603 protein" /label=ORF603 /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR" enhancer 893..1196 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1202..1479 /label=chicken beta-actin promoter primer_bind 1759..1781 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1843..1862 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" primer_bind 1904..1923 /label=T7 /note="T7 promoter, forward primer" promoter 1904..1922 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 1923..1947 /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 1963..2072 /label=p10 promoter /note="baculovirus promoter for expression in insect cells" CDS 2093..2110 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 2201..3229 /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="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED GD" CDS 3236..3259 /codon_start=1 /product="8xHis affinity tag" /label=8xHis /translation="HHHHHHHH" primer_bind complement(3349..3368) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 3414..3469 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(3468..3487) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" terminator 3557..3604 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" misc_recomb 3617..5075 /label=baculovirus recombination region (ORF1629) /note="contains part of ORF1629" 3'UTR 3705..4077 /label=polyhedrin 3' UTR /note="3' UTR from the Autographa californica nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)" CDS complement(3734..5075) /codon_start=2 /product="baculovirus capsid-associated protein" /label=ORF1629 /note="required for viral replication" /translation="PSLRQSLYNTIAYIERLLNIGTVNDSEITMLIADFYDLYSNYNIE LPPPQALPRSRRPSVVQPAAPAPVPTIVREQTKPEQIIPAAPPPPPSPVPNIPAPPPPP PPSMSELPPAPPMPTEPQPAAPLDDRQQLLEAIRNEKNRTRLRPVKPKTAPETSTIVEV PTVLPKETFEPKPPSASPPPPPPPPPPPAPPAPPPMVDLSSAPPPPPLVDLPSEMLPPP APSLSNVLSELKSGTVRLKPAQKRPQSEIIPKSSTTNLIADVLADTINRRRVAMAKSSS EATSNDEGWDDDDNRPNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVE PLLQNKTQTNIDKARLLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDL IFKFRYKDAENHLIFALTYHPKDYKFNELLKYVQQLSVNQQRTESSA" rep_origin complement(5234..5822) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5314..5333) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5835..6695) /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 6458..6477 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" ORIGIN 1 tcctgcatct tttaatcaaa tcccaagatg tgtataaacc accaaactgc caaaaaatga 61 aaactgtcga caagctctgt ccgtttgctg gcaactgcaa gggtctcaat cctatttgta 121 attattgaat aataaaacaa ttataaatgt caaatttgtt ttttattaac gatacaaacc 181 aaacgcaaca agaacatttg tagtattatc tataattgaa aacgcgtagt tataatcgct 241 gaggtaatat ttaaaatcat tttcaaatga ttcacagtta atttgcgaca atataatttt 301 attttcacat aaactagacg ccttgtcgtc ttcttcttcg tattccttct ctttttcatt 361 tttctcttca taaaaattaa catagttatt atcgtatcca tatatgtatc tatcgtatag 421 agtaaatttt ttgttgtcat aaatatatat gtctttttta atggggtgta tagtaccgct 481 gcgcatagtt tttctgtaat ttacaacagt gctattttct ggtagttctt cggagtgtgt 541 tgctttaatt attaaattta tataatcaat gaatttggga tcgtcggttt tgtacaatat 601 gttgccggca tagtacgcag cttcttctag ttcaattaca ccatttttta gcagcaccgg 661 attaacataa ctttccaaaa tgttgtacga accgttaaac aaaaacagtt cacctccctt 721 ttctatacta ttgtctgcga gcagttgttt gttgttaaaa ataacagcca ttgtaatgag 781 acgcacaaac taatatcaca aactggaaat gtctatcaat atatagttgc tctagttatt 841 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat 901 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa 961 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg 1021 actatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc 1081 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct 1141 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatgcatg 1201 gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca 1261 attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg gggggggggg 1321 gggcgcgcgc caggcggggc ggggcggggc gaggggcggg gcggggcgag gcggagaggt 1381 gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg 1441 cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gacgctgcct 1501 tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc 1561 gttactccca caggtgagcg ggcgggacgg cccttctcct tcgggctgta attagcgctt 1621 ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg ggctccggga 1681 gggccctttg tgcgggggga gcggctcggg gctgtccgcg gggggacggc tgccttcggg 1741 ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg 1801 ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg 1861 tgctgtctca tcattttggc aaagaattgg atcggaccga aattaatacg actcactata 1921 ggggaattgt gagcggataa caattccccg gagttaatcc gggaccttta attcaaccca 1981 acacaatata ttatagttaa ataagaatta ttatcaaatc atttgtatat taattaaaat 2041 actatactgt aaattacatt ttatttacaa tcaaaggaga tataccatgg gccatcacca 2101 tcaccatcac ggcatgggcg ctgcaggtcg caagaaacgt cgccaacgtc gccgtccgcc 2161 tgcaggcact agtgtaagct tgaagaagaa gaggaaggtg tccaatttac tgaccgtaca 2221 ccaaaatttg cctgcattac cggtcgatgc aacgagtgat gaggttcgca agaacctgat 2281 ggacatgttc agggatcgcc aggcgttttc tgagcatacc tggaaaatgc ttctgtccgt 2341 ttgccggtcg tgggcggcat ggtgcaagtt gaataaccgg aaatggtttc ccgcagaacc 2401 tgaagatgtt cgcgattatc ttctatatct tcaggcgcgc ggtctggcag taaaaactat 2461 ccagcaacat ttgggccagc taaacatgct tcatcgtcgg tccgggctgc cacgaccaag 2521 tgacagcaat gctgtttcac tggttatgcg gcggatccga aaagaaaacg ttgatgccgg 2581 tgaacgtgca aaacaggctc tagcgttcga acgcactgat ttcgaccagg ttcgttcact 2641 catggaaaat agcgatcgct gccaggatat acgtaatctg gcatttctgg ggattgctta 2701 taacaccctg ttacgtatag ccgaaattgc caggatcagg gttaaagata tctcacgtac 2761 tgacggtggg agaatgttaa tccatattgg cagaacgaaa acgctggtta gcaccgcagg 2821 tgtagagaag gcacttagcc tgggggtaac taaactggtc gagcgatgga tttccgtctc 2881 tggtgtagct gatgatccga ataactacct gttttgccgg gtcagaaaaa atggtgttgc 2941 cgcgccatct gccaccagcc agctatcaac tcgcgccctg gaagggattt ttgaagcaac 3001 tcatcgattg atttacggcg ctaaggatga ctctggtcag agatacctgg cctggtctgg 3061 acacagtgcc cgtgtcggag ccgcgcgaga tatggcccgc gctggagttt caataccgga 3121 gatcatgcaa gctggtggct ggaccaatgt aaatattgtc atgaactata tccgtaacct 3181 ggatagtgaa acaggggcaa tggtgcgcct gctggaagat ggcgattagc tcgagcacca 3241 ccatcaccat caccatcact aagtgattaa cctcaggtgc aggctgccta tcagaaggtg 3301 gtggctggtg tggccaatgc cctggctcac aaataccact gagatcgatc tttttccctc 3361 tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag 3421 gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac 3481 atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac 3541 atatgcccat atgtaactag cataacccct tggggcctct aaacgggtct tgaggggttt 3601 tttgctgaaa gcatgcggag gaaattctcc ttgaagtttc cctggtgttc aaagtaaagg 3661 agtttgcacc agacgcacct ctgttcactg gtccggcgta ttaaaacacg atacattgtt 3721 attagtacat ttattaagcg ctagattctg tgcgttgttg atttacagac aattgttgta 3781 cgtattttaa taattcatta aatttataat ctttagggtg gtatgttaga gcgaaaatca 3841 aatgattttc agcgtcttta tatctgaatt taaatattaa atcctcaata gatttgtaaa 3901 ataggtttcg attagtttca aacaagggtt gtttttccga accgatggct ggactatcta 3961 atggattttc gctcaacgcc acaaaacttg ccaaatcttg tagcagcaat ctagctttgt 4021 cgatattcgt ttgtgttttg ttttgtaata aaggttcgac gtcgttcaaa atattatgcg 4081 cttttgtatt tctttcatca ctgtcgttag tgtacaattg actcgacgta aacacgttaa 4141 atagagcttg gacatattta acatcgggcg tgttagcttt attaggccga ttatcgtcgt 4201 cgtcccaacc ctcgtcgtta gaagttgctt ccgaagacga ttttgccata gccacacgac 4261 gcctattaat tgtgtcggct aacacgtccg cgatcaaatt tgtagttgag ctttttggaa 4321 ttatttctga ttgcgggcgt ttttgggcgg gtttcaatct aactgtgccc gattttaatt 4381 cagacaacac gttagaaagc gatggtgcag gcggtggtaa catttcagac ggcaaatcta 4441 ctaatggcgg cggtggtgga gctgatgata aatctaccat cggtggaggc gcaggcgggg 4501 ctggcggcgg aggcggaggc ggaggtggtg gcggtgatgc agacggcggt ttaggctcaa 4561 atgtctcttt aggcaacaca gtcggcacct caactattgt actggtttcg ggcgccgttt 4621 ttggtttgac cggtctgaga cgagtgcgat ttttttcgtt tctaatagct tccaacaatt 4681 gttgtctgtc gtctaaaggt gcagcgggtt gaggttccgt cggcattggt ggagcgggcg 4741 gcaattcaga catcgatggt ggtggtggtg gtggaggcgc tggaatgtta ggcacgggag 4801 aaggtggtgg cggcggtgcc gccggtataa tttgttctgg tttagtttgt tcgcgcacga 4861 ttgtgggcac cggcgcaggc gccgctggct gcacaacgga aggtcgtctg cttcgaggca 4921 gcgcttgggg tggtggcaat tcaatattat aattggaata caaatcgtaa aaatctgcta 4981 taagcattgt aatttcgcta tcgtttaccg tgccgatatt taacaaccgc tcaatgtaag 5041 caattgtatt gtaaagagat tgtctcaagc tcggaacgct gcgctcggtc gttcggctgc 5101 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 5161 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 5221 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 5281 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 5341 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 5401 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt 5461 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 5521 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 5581 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 5641 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc 5701 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 5761 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 5821 aagaagatcc tttgttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 5881 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 5941 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 6001 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 6061 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 6121 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 6181 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 6241 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 6301 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 6361 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 6421 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 6481 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 6541 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 6601 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 6661 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 6721 agcatttatc agggttattg tctcatgtcc gcgcgtt //