LOCUS       sequence-271719-        6852 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 6852)
  TITLE     SGC+year
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 6852)
  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: 
            "unpublished"
COMMENT     Sequence Label: TXNRD1
FEATURES             Location/Qualifiers
     source          1..6852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     4..23
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     107..126
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        107..125
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    126..150
                     /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             165..187
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             198..215
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             240..260
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1823..1840
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1893..1911)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1907..1954
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1991..2446
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2078..2097)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2288..2309
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2538..3353)
                     /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             complement(2538..3353)
                     /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     3261..3280
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3345..3364
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3475..4063
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3964..3983
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4217..4234
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4249..4388
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4314..4333)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4474..4496
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4490..4681)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5490..6572)
                     /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     6534..6553
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6573..6650)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
       61 cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat acgactcact
      121 ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt taactttaag
      181 aaggagatat acatatgcac catcatcatc atcattcttc tggtgtagat ctgggtaccg
      241 agaacctgta cttccaatcc atgaacggcc ctgaagatct tcccaagtcc tatgactatg
      301 accttatcat cattggaggt ggctcaggag gtctggcagc tgctaaggag gcagcccaat
      361 atggcaagaa ggtgatggtc ctggactttg tcactcccac ccctcttgga actagatggg
      421 gtcttggagg aacatgtgtg aatgtgggtt gcatacctaa aaaactgatg catcaagcag
      481 ctttgttagg acaagccctg caagactctc gaaattatgg atggaaagtc gaggagacag
      541 ttaagcatga ttgggacaga atgatagaag ctgtacagaa tcacattggc tctttgaatt
      601 ggggctaccg agtagctctg cgggagaaaa aagtcgtcta tgagaatgct tatgggcaat
      661 ttattggtcc tcacaggatt aaggcaacaa ataataaagg caaagaaaaa atttattcag
      721 cagagagatt tctcattgcc actggtgaaa gaccacgtta cttgggcatc cctggtgaca
      781 aagaatactg catcagcagt gatgatcttt tctccttgcc ttactgcccg ggtaagaccc
      841 tggttgttgg agcatcctat gtcgctttgg agtgcgctgg atttcttgct ggtattggtt
      901 tagacgtcac tgttatggtt aggtccattc ttcttagagg atttgaccag gacatggcca
      961 acaaaattgg tgaacacatg gaagaacatg gcatcaagtt tataagacag ttcgtaccaa
     1021 ttaaagttga acaaattgaa gcagggacac caggccgact cagagtagta gctcagtcca
     1081 ccaatagtga ggaaatcatt gaaggagaat ataatacggt gatgctggca ataggaagag
     1141 atgcttgcac aagaaaaatt ggcttagaaa ccgtaggggt gaagataaat gaaaagactg
     1201 gaaaaatacc tgtcacagat gaagaacaga ccaatgtgcc ttacatctat gccattggcg
     1261 atatattgga ggataaggtg gagctcaccc cagttgcaat ccaggcagga agattgctgg
     1321 ctcagaggct ctatgcaggt tccactgtca agtgtgacta tgaaaatgtt ccaaccactg
     1381 tatttactcc tttggaatat ggtgcttgtg gcctttctga ggagaaagct gtggagaagt
     1441 ttggggaaga aaatattgag gtttaccata gttacttttg gccattggaa tggacgattc
     1501 cgtcaagaga taacaacaaa tgttatgcaa aaataatctg taatactaaa gacaatgaac
     1561 gtgttgtggg ctttcacgta ctgggtccaa atgctggaga agttacacaa ggctttgcag
     1621 ctgcgctcaa atgtggactg accaaaaagc agctggacag cacaattgga atccaccctg
     1681 tctgtgcaga ggtattcaca acattgtctg tgaccaagcg ctctggggca agcatcctcc
     1741 aggctggctg cggtggttaa cagtaaaggt ggatacggat ccgaattcga gctccgtcga
     1801 caagcttgcg gccgcactcg agcaccacca ccaccaccac tgagatccgg ctgctaacaa
     1861 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     1921 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatt
     1981 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2041 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2101 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2161 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2221 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2281 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2341 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2401 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     2461 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2521 cgctcatgaa ttaattctta gaaaaactca tcgagcatca aatgaaactg caatttattc
     2581 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
     2641 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt
     2701 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
     2761 tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag
     2821 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
     2881 ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa
     2941 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt
     3001 tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg
     3061 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
     3121 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
     3181 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc
     3241 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
     3301 ttggaattta atcgcggcct agagcaagac gtttcccgtt gaatatggct cataacaccc
     3361 cttgtattac tgtttatgta agcagacagt tttattgttc atgaccaaaa tcccttaacg
     3421 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3481 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3541 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3601 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     3661 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3721 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3781 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3841 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3901 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3961 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4021 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4081 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     4141 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     4201 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     4261 ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc
     4321 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
     4381 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     4441 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     4501 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     4561 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     4621 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     4681 ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag
     4741 agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg
     4801 agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat
     4861 gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga
     4921 tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa
     4981 cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt
     5041 cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc
     5101 agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggg gccgccatgc
     5161 cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt
     5221 gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc
     5281 agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt
     5341 gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga
     5401 aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag
     5461 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5521 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5581 gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc
     5641 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc
     5701 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc
     5761 actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc
     5821 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt
     5881 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc
     5941 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca
     6001 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc
     6061 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca
     6121 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc
     6181 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc
     6241 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca
     6301 cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc
     6361 agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg
     6421 cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca
     6481 gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac
     6541 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc
     6601 gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg
     6661 acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt
     6721 gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc
     6781 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc
     6841 ccgatcttcc cc
//
