LOCUS       sequence-175211-        6001 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of SELENOM CXXC for protein production.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6001)
  AUTHORS   Liu J, Chen Q, Rozovsky S
  TITLE     Utilizing Selenocysteine for Expressed Protein Ligation and 
            Bioconjugations.
  JOURNAL   J Am Chem Soc. 2017 Feb 10. doi: 10.1021/jacs.6b10991.
  PUBMED    28186733
REFERENCE   2  (bases 1 to 6001)
  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 Am Chem 
            Soc. 2017 Feb 10. doi: 10.1021/jacs.6b10991."
COMMENT     Sequence Label: SELENOM CXXC
FEATURES             Location/Qualifiers
     source          1..6001
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..29)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..53
                     /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     42..64
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             132..149
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     1215..1238
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1254..1274
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     terminator      1682..1768
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1860..1887
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1915..2005
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2006..2074,2075..2866)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAWQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2224..2243)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2954..3542
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3443..3462
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3696..3713
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3896..3918
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3912..4103)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        4599..4676
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             4677..5759
                     /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(4696..4715)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag ccagtccgtt taggtgtttt cacgagcaat tgaccaacaa ggaccataga
      121 ttatgaaaat ccatcaccat caccatcacg aagaaggtaa actggtaatc tggattaacg
      181 gcgataaagg ctataacggt ctcgctgaag tcggtaagaa attcgagaaa gataccggaa
      241 ttaaagtcac cgttgagcat ccggataaac tggaagagaa attcccacag gttgcggcaa
      301 ctggcgatgg ccctgacatt atcttctggg cacacgaccg ctttggtggc tacgctcaat
      361 ctggcctgtt ggctgaaatc accccggaca aagcgttcca ggacaagctg tatccgttta
      421 cctgggatgc cgtacgttac aacggcaagc tgattgctta cccgatcgct gttgaagcgt
      481 tatcgctgat ttataacaaa gatctgctga cgaacccgcc aaaaacctgg gaagagatcc
      541 cggcgctgga taaagaactg aaagcgaaag gtaagagcgc gctgatgttc aacctgcaag
      601 aaccgtactt cacctggccg ctgattgctg ctgacggggg ttatgcgttc aagtatgaaa
      661 acggcaagta cgacattaaa gacgtgggcg tggataacgc tggcgcgaaa gcgggtctga
      721 ccttcctggt tgacctgatt aaaaacaaac acatgaatgc agacaccgat tactccatcg
      781 cagaagctgc ctttaataaa ggcgaaacag cgatgaccat caacggcccg tgggcatggt
      841 ccaacatcga caccagcaaa gtgaattatg gtgtaacggt actgccgacc ttcaagggtc
      901 aaccatccaa accgttcgtt ggcgtgctga gcgcaggtat taacgccgcc agtccgaaca
      961 aagagctggc aaaagagttc ctcgaaaact atctgctgac tgatgaaggt ctggaagcgg
     1021 ttaataaaga caaaccgctg ggtgccgtag cgctgaagtc ttacgaggaa gagttggtga
     1081 aagatccgcg tattgccgcc actatggaaa acgcccagaa aggtgaaatc atgccgaaca
     1141 tcccgcagat gtccgctttc tggtatgccg tgcgtactgc ggtgatcaac gccgccagcg
     1201 gtcgtcagac tgtcgatgaa gccctgaaag acgcgcagac taattcgagc tctgaaaacc
     1261 tgtattttca gtccatgggc accgcttacc gcccggattg gaaccgtctg agtggcctga
     1321 cccgtgcgcg tgtggaaacg tgcggtggtt gtcagctgaa ccgcctgaaa gaagtgaagg
     1381 cttttgttac ccaagatatt ccgttctatc ataatctggt tatgaaacac ctgccgggcg
     1441 cagacccgga actggtcctg ctgggtcgtc gctacgaaga actggaacgt attccgctga
     1501 gcgaaatgac ccgcgaagaa atcaatgcgc tggttcagga actgggtttt tatcgtaaag
     1561 cagcaccgga tgcacaagtc ccgccggaat acgtgtgggc accggctaag ccgccggaag
     1621 aaacgtctga tcatgccgac ctgtaaggat ccgaattccc tgcaggtaat taaataagct
     1681 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     1741 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca
     1801 acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca
     1861 gaaggccatc ctgacggatg gcctttttgc gtttctacaa actctttcgg tccgttgttt
     1921 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     1981 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     2041 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     2101 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     2161 taagatcctt gagagttttc gccccgaaga acgtttccca atgatgagca cttttaaagt
     2221 tctgctatgt ggcgcggtat tatcccgtgt tgacgcctgg caagagcaac tcggtcgccg
     2281 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     2341 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     2401 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     2461 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     2521 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     2581 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     2641 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     2701 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     2761 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     2821 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     2881 ttactcatat atactttaga ttgatttcct taggactgag cgtcaacccc gtagaaaaga
     2941 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3001 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3061 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     3121 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3181 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3241 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3301 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3361 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3421 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3481 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3541 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3601 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     3661 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     3721 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
     3781 aaggtgcact gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     3841 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     3901 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
     3961 agctcatcag cgtggtcgtg cagcgattca cagatgtctg cctgttcatc cgcgtccagc
     4021 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
     4081 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg
     4141 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat
     4201 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag
     4261 agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag
     4321 ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc
     4381 cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc
     4441 gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc
     4501 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg
     4561 cgcacccgtg gccaggaccc aacgctgccc gaaattccga caccatcgaa tggtgcaaaa
     4621 cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga
     4681 aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc
     4741 gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga
     4801 tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt
     4861 tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg
     4921 cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa
     4981 gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc
     5041 tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta
     5101 atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt attattttct
     5161 cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa
     5221 tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc
     5281 ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg
     5341 ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga
     5401 tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc
     5461 tgcgcgttgg tgcggatatt tcggtagtgg gatacgacga taccgaagac agctcatgtt
     5521 atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg
     5581 accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct
     5641 cactggtgaa aagaaaaacc accctggcgc ccaatacaca aaccgcctct ccccgcgcgt
     5701 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     5761 cgcaacgcaa ttaatgtaag ttagctcact cattaggcac aattctcatg tttgacagct
     5821 tatcatcgac tgcacggtgc accaatgctt ctggcgtcag gcagccatcg gaagctgtgg
     5881 tatggctgtg caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg cactcccgtt
     5941 ctggataatg ttttttgcgc cgacatcata acggttctgg caaatattct gaaatgagct
     6001 g
//
