LOCUS       Exported                4550 bp ds-DNA     circular SYN 17-JAN-2020
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4550)
  AUTHORS   Twistbioscience
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jul 22, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     Insert is cloned directly after 3' C in the minimal Kozac site CACC 
            Insert GAC
FEATURES             Location/Qualifiers
     source          1..4550
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     83..99
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    152..251
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      270..279
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             276..2237
                     /codon_start=1
                     /label=lexA Transactivator
                     /label=lexA\Transactivator
                     /note="/vntifkey=21"
                     /translation="MAPKKKRKMKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGF
                     RSPNAAEEHLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAAGEPSSAGTEFPG
                     VDQKKFNKVRVVRALDAVALPQPVGVPNESQALSQRFTFSPGQDIQLIPPLINLLMSIE
                     PDVIYAGHDNTKPDTSSSLLTSLNQLGERQLLSVVKWSKSLPGFRNLHIDDQITLIQYS
                     WMSLMVFGLGWRSYKHVSGQMLYFAPDLILNEQRMKESSFYSLCLTMWQIPQEFVKLQV
                     SQEEFLCMKVLLLLNTIPLEGLRSQTQFEEMRSSYIRELIKAIGLRQKGVVSSSQRFYQ
                     LTKLLDNLHDLVKQLHLYCLNTFIQSRALSVEFPEMMSEVIAGSTPMEFQYLPDTDDRH
                     RIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSL
                     STINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPVPVLA
                     PGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSE
                     FQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDE
                     DFSSIADMDFSALLSQISS"
     CDS             276..299
                     /codon_start=1
                     /label=SV40 NLS
                     /translation="MAPKKKRK"
     CDS             300..551
                     /codon_start=1
                     /gene="lexA (truncated)"
                     /product="DNA-binding domain of E. coli LexA"
                     /label=LexA repressor DBD
                     /note="binds the 16-bp palindromic sequence 
                     5'-CTGTATATATATACAG-3'"
                     /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAA"
     primer_bind     complement(558..581)
                     /label=Frag 1 REV
     primer_bind     564..582
                     /label=oJDW XXX (lexA Seq FWD 1)
     misc_feature    573..596
                     /label=synthetic filler
     primer_bind     576..605
                     /label=middle frag FWD
     CDS             597..1865
                     /codon_start=1
                     /product="Human progesterone receptor binding domain"
                     /label=Progesterone Binding Domain
                     /translation="KKFNKVRVVRALDAVALPQPVGVPNESQALSQRFTFSPGQDIQLI
                     PPLINLLMSIEPDVIYAGHDNTKPDTSSSLLTSLNQLGERQLLSVVKWSKSLPGFRNLH
                     IDDQITLIQYSWMSLMVFGLGWRSYKHVSGQMLYFAPDLILNEQRMKESSFYSLCLTMW
                     QIPQEFVKLQVSQEEFLCMKVLLLLNTIPLEGLRSQTQFEEMRSSYIRELIKAIGLRQK
                     GVVSSSQRFYQLTKLLDNLHDLVKQLHLYCLNTFIQSRALSVEFPEMMSEVIAGSTPME
                     FQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKP
                     APQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSAL
                     AQAPAPVPVLAPGPPQAVAPPAPK"
     primer_bind     1080..1098
                     /label=oJDW XXX (lexA Seq FWD 2)
     primer_bind     1524..1546
                     /label=oJDW XXX (lexA Seq FWD 3)
     CDS             1866..2237
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA 
                     (NFkB), also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) Activation Domain
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALLSQISS"
     primer_bind     2005..2025
                     /label=oJDW XXX (lexA Seq FWD 4)
     CDS             2253..2372
                     /codon_start=1
                     /gene="Mus musculus ornithine decarboxylase gene 
                     destablization sequence.
                     
                     http://www.jbc.org/content/278/14/12135.abstract
                     "
                     /product="COOH 37 amino acids of mODC"
                     /label=mODC DEST
                     /note="mm Odc1 destablization domain"
                     /translation="SHGFPPAVAAQDDGTLPMSCAQESGMDRHPAACASARINV"
     protein_bind    complement(2437..2536)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2593..2609)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(2781..3369)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3505..4320)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     terminator      4458..4504
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 aacaaaaccc atcgtacggc caaggaagtc tccaataact gtgatccacc acaagcgcca
       61 gggttttccc agtcacgacg ttgtaaaacg acggccagtc atgcataatc cgcacgcatc
      121 tggaataagg aagtgccatt ccgcctgacc tcaaataatg attttatttt gactgatagt
      181 gacctgttcg ttgcaacaga ttgatgagca atgctttttt ataatgccaa ctttgtacaa
      241 aaaagcaggc ttacgtaact agtggatccg ccaccatggc acccaagaag aagaggaaga
      301 tgaaagcgtt aacggccagg caacaagagg tgtttgatct catccgtgat cacatcagcc
      361 agacaggtat gccgccgacg cgtgcggaaa tcgcgcagcg tttggggttc cgttccccaa
      421 acgcggctga agaacatctg aaggcgctgg cacgcaaagg cgttattgaa attgtttccg
      481 gcgcatcacg cgggattcgt ctgttgcagg aagaggaaga agggttgccg ctggtaggtc
      541 gtgtggctgc cggtgaaccg tcgagcgccg gtaccgaatt cccgggtgtc gaccagaaaa
      601 agttcaataa agtcagagtt gtgagagcac tggatgctgt tgctctccca cagccagtgg
      661 gcgttccaaa tgaaagccaa gccctaagcc agagattcac tttttcacca ggtcaagaca
      721 tacagttgat tccaccactg atcaacctgt taatgagcat tgaaccagat gtgatctatg
      781 caggacatga caacacaaaa cctgacacct ccagttcttt gctgacaagt cttaatcaac
      841 taggcgagag gcaacttctt tcagtagtca agtggtctaa atcattgcca ggttttcgaa
      901 acttacatat tgatgaccag ataactctca ttcagtattc ttggatgagc ttaatggtgt
      961 ttggtctagg atggagatcc tacaaacacg tcagtgggca gatgctgtat tttgcacctg
     1021 atctaatact aaatgaacag cggatgaaag aatcatcatt ctattcatta tgccttacca
     1081 tgtggcagat cccacaggag tttgtcaagc ttcaagttag ccaagaagag ttcctctgta
     1141 tgaaagtatt gttacttctt aatacaattc ctttggaagg gctacgaagt caaacccagt
     1201 ttgaggagat gaggtcaagc tacattagag agctcatcaa ggcaattggt ttgaggcaaa
     1261 aaggagttgt gtcgagctca cagcgtttct atcaacttac aaaacttctt gataacttgc
     1321 atgatcttgt caaacaactt catctgtact gcttgaatac atttatccag tcccgggcac
     1381 tgagtgttga atttccagaa atgatgtctg aagttattgc tgggtcgacg cccatggaat
     1441 ttcagtacct gccagataca gacgatcgtc accggattga ggagaaacgt aaaaggacat
     1501 atgagacctt caagagcatc atgaagaaga gtcctttcag cggacccacc gacccccggc
     1561 ctccacctcg acgcattgct gtgccttccc gcagctcagc ttctgtcccc aagccagcac
     1621 cccagcccta tccctttacg tcatccctga gcaccatcaa ctatgatgag tttcccacca
     1681 tggtgtttcc ttctgggcag atcagccagg cctcggcctt ggccccggcc cctccccaag
     1741 tcctgcccca ggctccagcc cctgcccctg ctccagccat ggtatcagct ctggcccagg
     1801 ccccagcccc tgtcccagtc ctagccccag gccctcctca ggctgtggcc ccacctgccc
     1861 ccaagcccac ccaggctggg gaaggaacgc tgtcagaggc cctgctgcag ctgcagtttg
     1921 atgatgaaga cctgggggcc ttgcttggca acagcacaga cccagctgtg ttcacagacc
     1981 tggcatccgt cgacaactcc gagtttcagc agctgctgaa ccagggcata cctgtggccc
     2041 cccacacaac tgagcccatg ctgatggagt accctgaggc tataactcgc ctagtgacag
     2101 gggcccagag gccccccgac ccagctcctg ctccactggg ggccccgggg ctccccaatg
     2161 gcctcctttc aggagatgaa gacttctcct ccattgcgga catggacttc tcagccctgc
     2221 tgagtcagat cagctccaga tctcgagata tcagccatgg cttcccgccg gcggtggcgg
     2281 cgcaggatga tggcacgctg cccatgtctt gtgcccagga gagcgggatg gaccgtcacc
     2341 ctgcagcctg tgcttctgct aggatcaatg tgtagtaaat cgatgatcct ccacgtagcg
     2401 gccgcgttta aacgctagca actctagatt aattaaaccc agctttcttg tacaaagttg
     2461 gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa
     2521 ataaaatcat tatttgaggc taggtggagg ctcagtgatg ataagtctgc gatggtggat
     2581 gcatgtgtca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca gagggcacaa
     2641 tcctattccg cgctatccga caatctccaa gacattaggt ggagttcagt tcggcgtatg
     2701 gcatatgtcg ctggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2761 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2821 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2881 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2941 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3001 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3061 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3121 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3181 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3241 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3301 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3361 ggatctcaag aagatccttt gatcttttct acggggtctg acgctctatt caacaaagcc
     3421 gccgtcccgt caagtcagcg taaatgggta gggggcttca aatcgtccgc tctgccagtg
     3481 ttacaaccaa ttaacaaatt ctgattagaa aaactcatcg agcatcaaat gaaactgcaa
     3541 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
     3601 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
     3661 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
     3721 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagct tatgcatttc
     3781 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac
     3841 caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa
     3901 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac
     3961 aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat
     4021 cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag
     4081 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac
     4141 gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata
     4201 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc
     4261 atccatgttg gaatttaatc gcggcctcga gcaagacgtt tcccgttgaa tatggctcat
     4321 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt
     4381 tttatcttgt gcaatgtaac atcagagatt ttgagacaca acgtggcttt cccccgccgc
     4441 tctagaacta gtggatccaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     4501 ttatctgttg tttgtcgcat tatacgagac gtccaggttg ggatacctga
//
