LOCUS       sequence-248528-        5607 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  coding sequence for synthetic zinc finger ZF-BCB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5607)
  AUTHORS   Swank Z, Laohakunakorn N, Maerkl SJ
  TITLE     Cell-free gene-regulatory network engineering with synthetic 
            transcription factors.
  JOURNAL   Proc Natl Acad Sci U S A. 2019 Mar 26;116(13):5892-5901. doi: 
            10.1073/pnas.1816591116. Epub 2019 Mar 8.
  PUBMED    30850530
REFERENCE   2  (bases 1 to 5607)
  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"; doi: 
            "10.1073/pnas.1816591116"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2019-03-26- 26"; volume: "116"; issue: "13"; pages: 
            "5892-5901"
COMMENT     Sequence Label: pZF-BCB
FEATURES             Location/Qualifiers
     source          1..5607
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(324..342)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      338..385
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      422..877
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(509..528)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     719..740
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        903..1007
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1008..1076,1077..1868)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1226..1245)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2039..2627
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2528..2547
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2781..2798
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2813..2952
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2878..2897)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3038..3060
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3054..3245)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4054..5136)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5098..5117
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5137..5214)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5420..5439
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5527..5546
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5527..5545
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5546..5570
                     /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             5585..5607
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 tatacatatg gaacgtccgt acgcttgccc ggttgaatct tgcgaccgtc gtttctctcg
       61 taacttcatc ctgcagcgtc atattagaat tcatactgga caaaaaccat tccaatgtag
      121 aatttgtatg agaaatttct ctcagcgttc ttctctggtt cgtcacatcc gtacccacac
      181 cggtgaaaaa ccgttcgctt gcgacatctg cggtcgtaaa ttcgctcgtc acgaccagct
      241 gacccgtcac accaaaatcc acctgcgtca gtaagatccg gctgctaaca aagcccgaaa
      301 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
      361 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat tggcgaatgg
      421 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
      481 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      541 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      601 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      661 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
      721 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
      781 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
      841 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
      901 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
      961 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     1021 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     1081 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     1141 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     1201 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     1261 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     1321 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     1381 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     1441 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     1501 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg
     1561 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     1621 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     1681 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     1741 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     1801 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     1861 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     1921 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     1981 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2041 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2101 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2161 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     2221 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2281 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2341 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2401 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     2461 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     2521 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     2581 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     2641 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     2701 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     2761 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     2821 ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac
     2881 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
     2941 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     3001 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     3061 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
     3121 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
     3181 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
     3241 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
     3301 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
     3361 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
     3421 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
     3481 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac
     3541 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag
     3601 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc
     3661 cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg tggggccgcc
     3721 atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag
     3781 gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg
     3841 ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg
     3901 agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac
     3961 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa
     4021 tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4081 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4141 gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac
     4201 cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa
     4261 atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta
     4321 tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc
     4381 gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag
     4441 catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat
     4501 cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga
     4561 gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg
     4621 ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg
     4681 gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc
     4741 atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt
     4801 gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct
     4861 ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag
     4921 ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc
     4981 cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt
     5041 cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc
     5101 atactctgcg acatcgtata acgttactgg tttcacattc accaccctga attgactctc
     5161 ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat
     5221 ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc
     5281 cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc
     5341 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
     5401 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
     5461 cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatcgat ctcgatcccg
     5521 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa
     5581 taattttgtt taactttaag aaggaga
//
