LOCUS       sequence-178339-        7045 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 7045)
  TITLE     SGC+year
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 7045)
  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            "unpublished"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: RECQL
FEATURES             Location/Qualifiers
     source          1..7045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     CDS             1798..1818
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1819..1836
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1837..1860
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1910..1927
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1980..1998)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1994..2041
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2078..2533
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2165..2184)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2375..2396
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2625..3440)
                     /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(2625..3440)
                     /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     3348..3367
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3432..3451
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3562..4150
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4051..4070
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4304..4321
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4336..4475
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4401..4420)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4561..4583
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4577..4768)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5577..6659)
                     /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     6621..6640
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6660..6737)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6943..6962
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaacct taagaaggag atatactatg tgtttagagg attctgatgc cggggcaagc
      121 aatgaatatg attcttcacc tgccgcttgg aataaagaag attttccatg gtctggtaaa
      181 gttaaagata ttctgcaaaa tgtctttaaa ctggaaaagt tcagaccact tcagcttgaa
      241 actattaacg taacaatggc tggaaaggag gtatttcttg ttatgcctac aggaggtgga
      301 aagagcttat gttaccagtt accagcatta tgttcagatg gttttacact cgtcatttgc
      361 ccattgatct ctcttatgga agaccaatta atggttttaa aacaattagg aatttcagca
      421 accatgttaa atgcttctag ttctaaggag catgttaaat gggttcatgc tgaaatggta
      481 aataaaaact ccgagttaaa gctgatttat gtgactccag agaaaattgc aaaaagcaaa
      541 atgtttatgt caagactaga gaaagcctat gaagcaagga gatttactcg aattgctgtg
      601 gatgaagttc actgctgtag tcagtgggga catgatttca gacctgatta taaggcactt
      661 ggtatcttaa agcggcagtt ccctaacgca tcactaattg ggctgactgc aactgcaaca
      721 aatcacgttt tgacggatgc tcagaaaatt ttgtgcattg aaaagtgttt tacttttaca
      781 gcttctttta ataggccaaa tctatattat gaggttcggc agaagccctc aaacactgaa
      841 gattttattg aggatattgt aaagctcatt aatgggagat acaaagggca atcaggaatc
      901 atatattgtt tttctcagaa agactctgaa caagttacgg ttagtttgca gaatctggga
      961 attcatgcag gtgcttacca tgccaatttg gagccagaag ataagaccac agttcataga
     1021 aaatggtcag ccaatgaaat tcaggtagta gtggcaactg ttgcatttgg tatgggaatt
     1081 gataagccag atgtgaggtt tgttatccat cattcaatga gtaaatccat ggaaaattat
     1141 taccaagaga gtggacgtgc aggtcgagat gacatgaaag cagactgtat tttgtactac
     1201 ggctttggag atatattcag aataagttca atggtggtga tggaaaatgt gggacagcag
     1261 aagctttatg agatggtatc atactgtcaa aacataagca aatgtcgtcg tgtgttgatg
     1321 gctcaacatt ttgatgaagt atggaactca gaagcatgta acaaaatgtg cgataactgc
     1381 tgtaaagaca gtgcatttga aagaaagaac ataacagagt actgcagaga tctaatcaag
     1441 atcctgaagc aggcagagga actgaatgaa aaactcactc cattgaaact gattgattct
     1501 tggatgggaa agggtgcagc aaaactgaga gtagcaggtg ttgtggctcc cacacttcct
     1561 cgtgaagatc tggagaagat tattgcacac tttctaatac agcagtatct taaagaagac
     1621 tacagtttta cagcttatgc taccatttcg tatttgaaaa taggacctaa agctaacctt
     1681 ctgaacaatg aggcacatgc tattactatg caagtgacaa agtccacgca gaactctttc
     1741 agggctgaat cgtctcaaac ttgtcattct gaacaaagtg ataaaaagat ggaggcagag
     1801 aacctctact tccaatcgca ccatcatcac caccatgatt acaaggatga cgacgataag
     1861 tgaggatccg aattcgagct ccgtcgacaa gcttgcggcc gcactcgagc accaccacca
     1921 ccaccactga gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
     1981 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     2041 gctgaaagga ggaactatat ccggattggc gaatgggacg cgccctgtag cggcgcatta
     2101 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2161 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2221 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2281 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2341 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2401 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     2461 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     2521 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     2581 ttttctaaat acattcaaat atgtatccgc tcatgaatta attcttagaa aaactcatcg
     2641 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa
     2701 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc
     2761 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg
     2821 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
     2881 ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
     2941 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga
     3001 aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg
     3061 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg
     3121 aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata
     3181 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca
     3241 tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg
     3301 ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat
     3361 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctaga gcaagacgtt
     3421 tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt
     3481 attgttcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3541 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3601 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3661 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     3721 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3781 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3841 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3901 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3961 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4021 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4081 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4141 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4201 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     4261 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     4321 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4381 ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
     4441 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
     4501 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     4561 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
     4621 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
     4681 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
     4741 taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca
     4801 tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg
     4861 aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg
     4921 accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc
     4981 cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg
     5041 acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc
     5101 aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat
     5161 tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga
     5221 tcatgcgcac ccgtggggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt
     5281 tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa
     5341 gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga
     5401 gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga
     5461 cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca
     5521 tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac
     5581 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5641 cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg
     5701 ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg
     5761 ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat
     5821 gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg
     5881 gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca
     5941 gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc
     6001 cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag
     6061 ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc
     6121 tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa
     6181 ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg
     6241 caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca
     6301 ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt
     6361 tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg
     6421 acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac
     6481 tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc
     6541 gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa
     6601 acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca
     6661 ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg
     6721 cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa
     6781 gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa
     6841 ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca
     6901 agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata
     6961 ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag
     7021 gatcgagatc tcgatcccgc gaaat
//
