LOCUS       sequence-248851-        5915 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  coding sequence for synthetic zinc finger ZF-BCB-PDZ.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5915)
  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 5915)
  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-PDZ
FEATURES             Location/Qualifiers
     source          1..5915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..25
                     /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             40..62
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(698..716)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      712..759
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      796..1251
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(883..902)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1093..1114
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1277..1381
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1382..1450,1451..2242)
                     /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(1600..1619)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2413..3001
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2902..2921
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3155..3172
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3187..3326
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3252..3271)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3412..3434
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3428..3619)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4428..5510)
                     /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     5472..5491
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5511..5588)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5794..5813
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     join(5897..5915,1)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5897..5915
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga
       61 gatatacata tggaacgtcc gtacgcttgc ccggttgaat cttgcgaccg tcgtttctct
      121 cgtaacttca tcctgcagcg tcatattaga attcatactg gacaaaaacc attccaatgt
      181 agaatttgta tgagaaattt ctctcagcgt tcttctctgg ttcgtcacat ccgtacccac
      241 accggtgaaa aaccgttcgc ttgcgacatc tgcggtcgta aattcgctcg tcacgaccag
      301 ctgacccgtc acaccaaaat ccacctgcgt cagaaagacg gtggcggcgg ttctccagcg
      361 ccagcgccac cggaagcgct gcagcgccgc cgcgtgaccg tgcgcaaagc ggatgcgggc
      421 ggcctgggca ttagcattaa aggcggccgc gaaaacaaaa tgccgattct gattagcaaa
      481 atttttaaag gcctggcggc ggatcagacc gaagcgctgt ttgtgggcga tgcgattctg
      541 agcgtgaacg gcgaagatct gagcagcgcg acccatgatg aagcggtgca ggcgctgaaa
      601 aaaaccggca aagaagtggt gctggaagtg aaatatatga aagaataaga tccggctgct
      661 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
      721 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
      781 ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
      841 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
      901 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
      961 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     1021 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     1081 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     1141 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     1201 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     1261 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     1321 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     1381 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     1441 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     1501 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     1561 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     1621 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     1681 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     1741 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     1801 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     1861 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     1921 gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     1981 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     2041 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     2101 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     2161 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     2221 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     2281 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     2341 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     2401 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2461 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2521 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     2581 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2641 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2701 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     2761 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     2821 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     2881 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     2941 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3001 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3061 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     3121 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     3181 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg
     3241 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     3301 cgctacgtga ctgggtcatg gctgcgcccc aacacccgcc aacacccgct gacgcgccct
     3361 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     3421 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     3481 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     3541 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     3601 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
     3661 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
     3721 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
     3781 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
     3841 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
     3901 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
     3961 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
     4021 cagtaaggca accccgccag cctaaccggg tcctcaacga caggagcacg atcatgcgca
     4081 cccgtggggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg
     4141 gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc
     4201 cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg
     4261 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
     4321 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag
     4381 atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt
     4441 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     4501 gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc
     4561 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc
     4621 cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct
     4681 tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta
     4741 atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg
     4801 atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct
     4861 tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga
     4921 cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc
     4981 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg
     5041 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct
     5101 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt
     5161 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc
     5221 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc
     5281 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc
     5341 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact
     5401 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga
     5461 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc
     5521 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg
     5581 atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag
     5641 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat agtgcatgca aggagatggc
     5701 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
     5761 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
     5821 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat
     5881 ctcgatcccg cgaaattaat acgactcact atagg
//
