LOCUS       sequence-193346-        5543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of mouse CBP ZZ domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5543)
  AUTHORS   Legge GB, Martinez-Yamout MA, Hambly DM, Trinh T, Lee BM, Dyson HJ, 
            Wright PE
  TITLE     ZZ domain of CBP: an unusual zinc finger fold in a protein 
            interaction module.
  JOURNAL   J Mol Biol. 2004 Oct 29;343(4):1081-93.
  PUBMED    15476823
REFERENCE   2  (bases 1 to 5543)
  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: "J Mol 
            Biol."; date: "2004-10-29"; volume: "343(4)"; pages: "1081-93"
COMMENT     Sequence Label: pET-ZZ(1700-1751)
FEATURES             Location/Qualifiers
     source          1..5543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      25..613
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     514..533
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     767..784
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    799..938
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(864..883)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1024..1046
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1040..1231)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2040..3122)
                     /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     3084..3103
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3123..3200)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3406..3425
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3509..3528
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3509..3527
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3528..3552
                     /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             3567..3589
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3783..3800
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3853..3871)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3867..3914
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3951..4406
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4038..4057)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4248..4269
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4432..4536
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4537..4605,4606..5397)
                     /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(4755..4774)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
       61 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
      121 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
      181 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
      241 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
      301 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
      361 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
      421 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
      481 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
      541 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
      601 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
      661 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
      721 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
      781 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
      841 acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
      901 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
      961 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     1021 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     1081 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg
     1141 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     1201 tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt
     1261 tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg
     1321 atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc
     1381 gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg
     1441 ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg
     1501 ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg
     1561 ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt
     1621 cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca
     1681 cgatcatgcg cacccgtggg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac
     1741 gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg
     1801 caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc
     1861 agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg
     1921 cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     1981 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
     2041 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     2101 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
     2161 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
     2221 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
     2281 catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc
     2341 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     2401 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     2461 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     2521 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     2581 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
     2641 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
     2701 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
     2761 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
     2821 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
     2881 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
     2941 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
     3001 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
     3061 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
     3121 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
     3181 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag
     3241 gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg
     3301 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
     3361 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
     3421 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
     3481 gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg
     3541 gataacaatt cccctctaga aataattttg tttaacttta agaaggagat atacatatgg
     3601 gccaggaccg ctttgtttat acctgcaatg agtgcaaaca ccatgtggaa acacgctggc
     3661 actgcactgt gtgtgaggac tatgaccttt gtatcaattg ctacaacaca aagagccaca
     3721 cccataagat ggtgaagtgg gggctaggcc tagattaata gaagcttgcg gccgcactcg
     3781 agcaccacca ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt
     3841 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
     3901 tgaggggttt tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg
     3961 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4021 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4081 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4141 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     4201 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4261 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4321 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     4381 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     4441 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4501 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4561 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4621 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4681 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     4741 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     4801 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     4861 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     4921 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     4981 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5041 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     5101 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5161 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5221 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     5281 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     5341 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     5401 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5461 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5521 tcgttccact gagcgtcaga ccc
//
