LOCUS       sequence-495011-        6302 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express DBVen1619-T7RNAP that dimerize with BCL2 when Venetoclax 
            present.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6302)
  AUTHORS   Lee P-W, Mottaghi SS, Lugnier MG, Maerkl SJ
  TITLE     A T7 RNAP regulatory toolbox for cell-free network engineering and 
            biosensing applications
  JOURNAL   bioRxiv 2025.06.18.660355
REFERENCE   2  (bases 1 to 6302)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2025.06.18.660355"
COMMENT     Sequence Label: pQE30-His-DBVen1619-T7RNAP
FEATURES             Location/Qualifiers
     source          1..6302
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(171..962,963..1031))
                     /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     794..813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1032..1136)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1204..1222
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1246..1260,1261..1266,1267..1283,1284..1290)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    1266..1282
                     /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)."
     protein_bind    1298..1314
                     /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)."
     RBS             1333..1344
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1354..1371
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      4286..4380
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             4424..5083
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4490..4509)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      5148..5234
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(5281..5303)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5389..5529
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5444..5463
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5544..5561)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5715..6302)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5795..5814)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1141 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     1201 aaaaataggc gtatcacgag gccctttcgt cttcacctcg agaaatcata aaaaatttat
     1261 ttgctttgtg agcggataac aattataata gattcaattg tgagcggata acaatttcac
     1321 acagaattca ttaaagagga gaaattaact atgcatcacc accatcatca catgcagtat
     1381 ttgctggttg tcaaaggtcc taacgtgact aaattcagat gggtagactc aagttctgaa
     1441 gcggagacgc ttgctcgtaa gattgcaaaa aagctgggct tagaggtcaa aagtgtagaa
     1501 aaaaagggca atgctgtgcg tgtcgaaata ggatccgggg gcgggggatc cggaggcggg
     1561 ggatccggtg gtgggggttc tatgaacacg attaacatcg ctaagaacga cttctctgac
     1621 atcgaactgg ctgctatccc gttcaacact ctggctgacc attacggtga gcgtttagct
     1681 cgcgaacagt tggcccttga gcatgagtct tacgagatgg gtgaagcacg cttccgcaag
     1741 atgtttgagc gtcaacttaa agctggtgag gttgcggata acgctgccgc caagcctctc
     1801 atcactaccc tactccctaa gatgattgca cgcatcaacg actggtttga ggaagtgaaa
     1861 gctaagcgcg gcaagcgccc gacagccttc cagttcctgc aagaaatcaa gccggaagcc
     1921 gtagcgtaca tcaccattaa gaccactctg gcttgcctaa ccagtgctga caatacaacc
     1981 gttcaggctg tagcaagcgc aatcggtcgg gccattgagg acgaggctcg cttcggtcgt
     2041 atccgtgacc ttgaagctaa gcacttcaag aaaaacgttg aggaacaact caacaagcgc
     2101 gtagggcacg tctacaagaa agcatttatg caagttgtcg aggctgacat gctctctaag
     2161 ggtctactcg gtggcgaggc gtggtcttcg tggcataagg aagactctat tcatgtagga
     2221 gtacgctgca tcgagatgct cattgagtca accggaatgg ttagcttaca ccgccaaaat
     2281 gctggcgtag taggtcaaga ctctgagact atcgaactcg cacctgaata cgctgaggct
     2341 atcgcaaccc gtgcaggtgc gctggctggc atctctccga tgttccaacc ttgcgtagtt
     2401 cctcctaagc cgtggactgg cattactggt ggtggctatt gggctaacgg tcgtcgtcct
     2461 ctggcgctgg tgcgtactca cagtaagaaa gcactgatgc gctacgaaga cgtttacatg
     2521 cctgaggtgt acaaagcgat taacattgcg caaaacaccg catggaaaat caacaagaaa
     2581 gtcctagcgg tcgccaacgt aatcaccaag tggaagcatt gtccggtcga ggacatccct
     2641 gcgattgagc gtgaagaact cccgatgaaa ccggaagaca tcgacatgaa tcctgaggct
     2701 ctcaccgcgt ggaaacgtgc tgccgctgct gtgtaccgca aggacaaggc tcgcaagtct
     2761 cgccgtatca gccttgagtt catgcttgag caagccaata agtttgctaa ccataaggcc
     2821 atctggttcc cttacaacat ggactggcgc ggtcgtgttt acgctgtgtc aatgttcaac
     2881 ccgcaaggta acgatatgac caaaggactg cttacgctgg cgaaaggtaa accaatcggt
     2941 aaggaaggtt actactggct gaaaatccac ggtgcaaact gtgcgggtgt cgataaggtt
     3001 ccgttccctg agcgcatcaa gttcattgag gaaaaccacg agaacatcat ggcttgcgct
     3061 aagtctccac tggagaacac ttggtgggct gagcaagatt ctccgttctg cttccttgcg
     3121 ttctgctttg agtacgctgg ggtacagcac cacggcctga gctataactg ctcccttccg
     3181 ctggcgtttg acgggtcttg ctctggcatc cagcacttct ccgcgatgct ccgagatgag
     3241 gtaggtggtc gcgcggttaa cttgcttcct agtgaaaccg ttcaggacat ctacgggatt
     3301 gttgctaaga aagtcaacga gattctacaa gcagacgcaa tcaatgggac cgataacgaa
     3361 gtagttaccg tgaccgatga gaacactggt gaaatctctg agaaagtcaa gctgggcact
     3421 aaggcactgg ctggtcaatg gctggcttac ggtgttactc gcagtgtgac taagcgttca
     3481 gtcatgacgc tggcttacgg gtccaaagag ttcggcttcc gtcaacaagt gctggaagat
     3541 accattcagc cagctattga ttccggcaag ggtctgatgt tcactcagcc gaatcaggct
     3601 gctggataca tggctaagct gatttgggaa tctgtgagcg tgacggtggt agctgcggtt
     3661 gaagcaatga actggcttaa gtctgctgct aagctgctgg ctgctgaggt caaagataag
     3721 aagactggag agattcttcg caagcgttgc gctgtgcatt gggtaactcc tgatggtttc
     3781 cctgtgtggc aggaatacaa gaagcctatt cagacgcgct tgaacctgat gttcctcggt
     3841 cagttccgct tacagcctac cattaacacc aacaaagata gcgagattga tgcacacaaa
     3901 caggagtctg gtatcgctcc taactttgta cacagccaag acggtagcca ccttcgtaag
     3961 actgtagtgt gggcacacga gaagtacgga atcgaatctt ttgcactgat tcacgactcc
     4021 ttcggtacca ttccggctga cgctgcgaac ctgttcaaag cagtgcgcga aactatggtt
     4081 gacacatatg agtcttgtga tgtactggct gatttctacg accagttcgc tgaccagttg
     4141 cacgagtctc aattggacaa aatgccagca cttccggcta aaggtaactt gaacctccgt
     4201 gacatcttag agtcggactt cgcgttcgcg taactgcagg catcgcgttc gcgtaactgc
     4261 agccaagctt aattagctga gcttggactc ctgttgatag atccagtaat gacctcagaa
     4321 ctccatctgg atttgttcag aacgctcggt tgccgccggg cgttttttat tggtgagaat
     4381 ccaagctagc ttggcgagat tttcaggagc taaggaagct aaaatggaga aaaaaatcac
     4441 tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg aggcatttca
     4501 gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg cctttttaaa
     4561 gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc ttgcccgcct
     4621 gatgaatgct catccggaat ttcgtatggc aatgaaagac ggtgagctgg tgatatggga
     4681 tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt catcgctctg
     4741 gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag atgtggcgtg
     4801 ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt ttttcgtctc
     4861 agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata tggacaactt
     4921 cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc
     4981 gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca gaatgcttaa
     5041 tgaattacaa cagtactgcg atgagtggca gggcggggcg taattttttt aaggcagtta
     5101 ttggtgccct taaacgcctg gggtaatgac tctctagctt gaggcatcaa ataaaacgaa
     5161 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     5221 tgagtaggac aaatccgccc tctagagctg cctcgcgcgt ttcggtgatg acggtgaaaa
     5281 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     5341 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac
     5401 ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt
     5461 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     5521 cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5581 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5641 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5701 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5761 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5821 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5881 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5941 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6001 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6061 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6121 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     6181 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6241 gctggtagcg gtggtttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6301 aa
//
