LOCUS Exported 5531 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pOPINF SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5531) AUTHORS Berrow NS, Alderton D, Sainsbury S, Nettleship J, Assenberg R, Rahman N, Stuart DI, Owens RJ TITLE A versatile ligation-independent cloning method suitable for high-throughput expression screening applications. JOURNAL Nucleic Acids Res. 2007 . 35(6):e45. PUBMED 17317681 REFERENCE 2 (bases 1 to 5531) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5531 /organism="synthetic DNA construct" /mol_type="other DNA" misc_recomb 181..1008 /label=baculovirus recombination region (lef2/ORF603) /note="contains ORF603 and part of lef2" CDS 181..305 /codon_start=3 /product="baculovirus late expression factor 2" /label=lef2 /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY" CDS complement(343..948) /codon_start=1 /product="baculovirus ORF603 protein" /label=ORF603 /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR" enhancer 1140..1443 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1449..1726 /label=chicken beta-actin promoter primer_bind 2005..2027 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 2089..2108 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" primer_bind 2150..2169 /label=T7 /note="T7 promoter, forward primer" promoter 2150..2168 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 2169..2193 /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 2209..2318 /label=p10 promoter /note="baculovirus promoter for expression in insect cells" CDS 2339..2356 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" protein_bind 2430..2451 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 2466..2496 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2504..2520 /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)." primer_bind 2509..2531 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 2528..2544 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 2528..2544 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 2540..2713 /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART DRPSQQLRSLNGE" primer_bind complement(2560..2577) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2560..2576) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2569..2591) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(2826..2845) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 2891..2946 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(2945..2964) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" terminator 3034..3081 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" misc_recomb 3094..3799 /label=baculovirus recombination region (ORF1629) /note="contains part of ORF1629" 3'UTR 3182..3554 /label=polyhedrin 3' UTR /note="3' UTR from the Autographa californica nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)" CDS complement(3211..3799) /codon_start=2 /product="baculovirus capsid-associated protein" /label=ORF1629 /note="required for viral replication" /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA" protein_bind 3808..3841 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." rep_origin complement(4009..4596) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4088..4107) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4609..5469) /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 5232..5251 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" ORIGIN 1 tcctgcatct tttaatcaaa tcccaagatg tgtataaacg cgccggtatg tacaggaaga 61 ggtttatact aaactgttac attgcaaacg tggtttcgtg tgccaagtgt gaaaaccgat 121 gtttaatcaa ggctctgacg catttctaca accacgactc caagtgtgtg ggtgaagtca 181 tgcatctttt aatcaaatcc caagatgtgt ataaaccacc aaactgccaa aaaatgaaaa 241 ctgtcgacaa gctctgtccg tttgctggca actgcaaggg tctcaatcct atttgtaatt 301 attgaataat aaaacaatta taaatgtcaa atttgttttt tattaacgat acaaaccaaa 361 cgcaacaaga acatttgtag tattatctat aattgaaaac gcgtagttat aatcgctgag 421 gtaatattta aaatcatttt caaatgattc acagttaatt tgcgacaata taattttatt 481 ttcacataaa ctagacgcct tgtcgtcttc ttcttcgtat tccttctctt tttcattttt 541 ctcttcataa aaattaacat agttattatc gtatccatat atgtatctat cgtatagagt 601 aaattttttg ttgtcataaa tatatatgtc ttttttaatg gggtgtatag taccgctgcg 661 catagttttt ctgtaattta caacagtgct attttctggt agttcttcgg agtgtgttgc 721 tttaattatt aaatttatat aatcaatgaa tttgggatcg tcggttttgt acaatatgtt 781 gccggcatag tacgcagctt cttctagttc aattacacca ttttttagca gcaccggatt 841 aacataactt tccaaaatgt tgtacgaacc gttaaacaaa aacagttcac ctcccttttc 901 tatactattg tctgcgagca gttgtttgtt gttaaaaata acagccattg taatgagacg 961 cacaaactaa tatcacaaac tggaaatgtc tatcaatata tagttgctga tggccggccg 1021 taatgagacg cacaaactaa tatcacaaac tggaaatgtc tatcaatata tagttgctct 1081 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc 1141 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg 1201 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa 1261 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca 1321 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac 1381 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc 1441 atgcatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc 1501 acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg 1561 gggggggggg cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg 1621 gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag 1681 gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg 1741 ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact 1801 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctccttcgg gctgtaatta 1861 gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct 1921 ccgggagggc cctttgtgcg gggggagcgg ctcggggctg tccgcggggg gacggctgcc 1981 ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag 2041 cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg 2101 ttattgtgct gtctcatcat tttggcaaag aattggatcg gaccgaaatt aatacgactc 2161 actatagggg aattgtgagc ggataacaat tccccggagt taatccggga cctttaattc 2221 aacccaacac aatatattat agttaaataa gaattattat caaatcattt gtatattaat 2281 taaaatacta tactgtaaat tacattttat ttacaatcaa aggagatata ccatggcaca 2341 ccatcaccac catcacagca gcggtctgga agttctgttt cagggtaccg tcgacccgac 2401 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc 2461 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa 2521 tttcacacag gaaacagcta tgaccatgat tacggattca ctggccgtcg ttttacaacg 2581 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt 2641 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag 2701 cctgaatggc gaatggcgcc aagctttcta gaccagtttg tgattaacct caggtgcagg 2761 ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag 2821 atcgatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg 2881 acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt 2941 ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg 3001 gtttagagtt tggcaacata tgcccatatg taactagcat aaccccttgg ggcctctaaa 3061 cgggtcttga ggggtttttt gctgaaagca tgcggaggaa attctccttg aagtttccct 3121 ggtgttcaaa gtaaaggagt ttgcaccaga cgcacctctg ttcactggtc cggcgtatta 3181 aaacacgata cattgttatt agtacattta ttaagcgcta gattctgtgc gttgttgatt 3241 tacagacaat tgttgtacgt attttaataa ttcattaaat ttataatctt tagggtggta 3301 tgttagagcg aaaatcaaat gattttcagc gtctttatat ctgaatttaa atattaaatc 3361 ctcaatagat ttgtaaaata ggtttcgatt agtttcaaac aagggttgtt tttccgaacc 3421 gatggctgga ctatctaatg gattttcgct caacgccaca aaacttgcca aatcttgtag 3481 cagcaatcta gctttgtcga tattcgtttg tgttttgttt tgtaataaag gttcgacgtc 3541 gttcaaaata ttatgcgctt ttgtatttct ttcatcactg tcgttagtgt acaattgact 3601 cgacgtaaac acgttaaata gagcttggac atatttaaca tcgggcgtgt tagctttatt 3661 aggccgatta tcgtcgtcgt cccaaccctc gtcgttagaa gttgcttccg aagacgattt 3721 tgccatagcc acacgacgcc tattaattgt gtcggctaac acgtccgcga tcaaatttgt 3781 agttgagctt tttggaattg cgatcgcata acttcgtata gcatacatta tacgaagtta 3841 taagctcgga acgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 3901 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 3961 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 4021 ccgccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 4081 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 4141 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 4201 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 4261 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 4321 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 4381 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 4441 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 4501 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 4561 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgtt accaatgctt 4621 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 4681 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 4741 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 4801 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 4861 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 4921 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 4981 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 5041 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 5101 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 5161 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 5221 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 5281 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 5341 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 5401 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 5461 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 5521 gtccgcgcgt t //