LOCUS       sequence-248915-        5607 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  coding sequence for synthetic zinc finger ZF-BCD.
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-BCD
FEATURES             Location/Qualifiers
     source          1..5607
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             15..37
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(361..379)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      375..422
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      459..914
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(546..565)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     756..777
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        940..1044
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1045..1113,1114..1905)
                     /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(1263..1282)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2076..2664
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2565..2584
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2818..2835
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2850..2989
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2915..2934)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3075..3097
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3091..3282)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4091..5173)
                     /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     5135..5154
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5174..5251)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5457..5476
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5564..5583
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5564..5582
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5583..5607
                     /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)."
ORIGIN
        1 cctctagaaa taattttgtt taactttaag aaggagatat acatatggaa cgtccgtacg
       61 cttgcccggt tgaatcttgc gaccgtcgtt tctctgacaa aaccaaactg cgtgttcata
      121 ttagaattca tactggacaa aaaccattcc aatgtagaat ttgtatgaga aatttctctc
      181 agcgttcttc tctggttcgt cacatccgta cccacaccgg tgaaaaaccg ttcgcttgcg
      241 acatctgcgg tcgtaaattc gctcgtcacg accagctgac ccgtcacacc aaaatccacc
      301 tgcgtcagta agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
      361 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
      421 tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt
      481 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
      541 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
      601 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
      661 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
      721 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
      781 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
      841 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
      901 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
      961 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     1021 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     1081 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     1141 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     1201 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     1261 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     1321 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     1381 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     1441 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     1501 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     1561 aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta
     1621 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     1681 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     1741 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     1801 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     1861 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     1921 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     1981 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     2041 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     2101 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     2161 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     2221 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     2281 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     2341 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     2401 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     2461 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     2521 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     2581 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     2641 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     2701 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     2761 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     2821 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     2881 tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     2941 taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc
     3001 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3061 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3121 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc
     3181 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
     3241 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg
     3301 gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg
     3361 ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt
     3421 atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac
     3481 agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat
     3541 ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat
     3601 tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg
     3661 tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa
     3721 cgacaggagc acgatcatgc gcacccgtgg ggccgccatg ccggcgataa tggcctgctt
     3781 ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat
     3841 tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc
     3901 gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt
     3961 cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
     4021 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat
     4081 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4141 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
     4201 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
     4261 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
     4321 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac
     4381 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
     4441 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
     4501 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
     4561 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
     4621 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
     4681 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
     4741 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
     4801 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
     4861 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
     4921 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
     4981 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
     5041 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
     5101 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
     5161 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     5221 cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac
     5281 tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg
     5341 aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata
     5401 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg
     5461 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
     5521 cgtccggcgt agaggatcga gatcgatctc gatcccgcga aattaatacg actcactata
     5581 ggggaattgt gagcggataa caattcc
//
