LOCUS       sequence-472908-        5468 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression for structure determination; may not be full 
            ORF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5468)
  TITLE     Structural Genomics Consortium Clones
REFERENCE   2  (bases 1 to 5468)
  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"
COMMENT     Sequence Label: CBX2
FEATURES             Location/Qualifiers
     source          1..5468
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1452..1643
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1637..1659)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1745..1884
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1800..1819
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1899..1916)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2070..2658)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2150..2169)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(2769..2788)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2780..3595
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             2780..3595
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2853..2872)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(3687..4142)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3824..3845)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4036..4055
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(4179..4226)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4222..4240
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4293..4310)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4502..4522)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(4535..4552)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             4562..4584
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4599..4623
                     /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)."
     primer_bind     complement(4623..4642)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4624..4642)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4726..4745)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        4951..5028
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5029..5468,1..643)
                     /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     complement(5048..5067)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac
       61 gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg
      121 cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg
      181 caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca
      241 acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga
      301 tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga
      361 tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac
      421 caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact
      481 ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa
      541 aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
      601 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
      661 taagttagct cactcattag gcaccgggat ctcgaccgat gcccttgaga gccttcaacc
      721 cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct
      781 tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg
      841 accgctttcg ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc
      901 acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc gagaagcagg
      961 ccattatcgc cggcatggcg gccccacggg tgcgcatgat cgtgctcctg tcgttgagga
     1021 cccggctagg ctggcggggt tgccttactg gttagcagaa tgaatcaccg atacgcgagc
     1081 gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct
     1141 tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc accattatgt
     1201 tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca tctgtattaa
     1261 cgaagcgctg gcattgaccc tgagtgattt ttctctggtc ccgccgcatc cataccgcca
     1321 gttgtttacc ctcacaacgt tccagtaacc gggcatgttc atcatcagta acccgtatcg
     1381 tgagcatcct ctctcgtttc atcggtatca ttacccccat gaacagaaat cccccttaca
     1441 cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc tttatcagaa
     1501 gccagacatt aacgcttctg gagaaactca acgagctgga cgcggatgaa caggcagaca
     1561 tctgtgaatc gcttcacgac cacgctgatg agctttaccg cagctgcctc gcgcgtttcg
     1621 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     1681 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     1741 ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc
     1801 ggcatcagag cagattgtac tgagagtgca ccattgcggt gtgaaatacc gcacagatgc
     1861 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
     1921 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     1981 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     2041 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2101 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2161 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2221 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2281 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2341 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2401 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2461 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     2521 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2581 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2641 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2701 aacgaaaact cacgttaagg gattttggtc atgaacaata aaactgtctg cttacataaa
     2761 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcttgct ctaggccgcg
     2821 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     2881 gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct
     2941 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     3001 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     3061 atggttactc accactgcga tccccgggaa aacagcattc caggtattag aagaatatcc
     3121 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     3181 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     3241 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     3301 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
     3361 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     3421 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     3481 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     3541 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaagaatt
     3601 aattcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     3661 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
     3721 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     3781 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     3841 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     3901 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     3961 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     4021 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     4081 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     4141 gtcccattcg ccaatccgga tatagttcct cctttcagca aaaaacccct caagacccgt
     4201 ttagaggccc caaggggtta tgctagttat tgctcagcgg tggcagcagc caactcagct
     4261 tcctttcggg ctttgttagc agccggatct cagtggtggt ggtggtggtg ctcgagtgcg
     4321 gccgcaagct tcgtcatcat tccttcttct ggaaggccag gagcagcctc gggtccagga
     4381 tgttctcctc cggctcccag ctgttatgtt tggaggacca gccgcgccac ttgaccaggt
     4441 actccagctt gcccttgcgg agccgcttgc tcaggatgca ctcggcggcg aagacctgct
     4501 cgccctggaa atacaagttt tctctgccgc tgctgtgatg atgatgatga tgcatggtat
     4561 atctccttct taaagttaaa caaaattatt tctagagggg aattgttatc cgctcacaat
     4621 tcccctatag tgagtcgtat taatttcgcg ggatcgagat ctcgatcctc tacgccggac
     4681 gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca
     4741 tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg
     4801 gtatggtggc aggccccgtg gccgggggac tgttgggcgc catctccttg catgcaccat
     4861 tccttgcggc ggcggtgctc aacggcctca acctactact gggctgcttc ctaatgcagg
     4921 agtcgcataa gggagagcgt cgagatcccg gacaccatcg aatggcgcaa aacctttcgc
     4981 ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta
     5041 acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg
     5101 aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag
     5161 ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt
     5221 ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa
     5281 tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc
     5341 gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt
     5401 aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg
     5461 gcgttatt
//
