LOCUS       sequence-361429-        5819 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Baculovirus expression for structure determination. May not contain 
            entire coding region of gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5819)
  TITLE     SGC Structures-2021-Q3 and Q4
REFERENCE   2  (bases 1 to 5819)
  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: RFXANK:JMC029-A02:C213521
FEATURES             Location/Qualifiers
     source          1..5819
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(278..1360)
                     /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     1322..1341
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1361..1438)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1644..1663
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1747..1766
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1747..1765
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1766..1790
                     /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             1805..1827
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1837..1854
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1867..1887
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2430..2447
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2500..2518)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2514..2561
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2598..3053
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2685..2704)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2895..2916
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3145..3960)
                     /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(3145..3960)
                     /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     3868..3887
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3952..3971
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4082..4670
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4571..4590
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4824..4841
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4856..4995
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4921..4940)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5081..5103
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5097..5288)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt
       61 cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc
      121 tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc
      181 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc
      241 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg
      301 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
      361 tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct
      421 tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc
      481 gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt
      541 cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca
      601 ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat
      661 tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg
      721 ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg
      781 ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca
      841 gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg
      901 tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa
      961 tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa
     1021 gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca
     1081 cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt
     1141 gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt
     1201 gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg
     1261 ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac
     1321 cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac
     1381 tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg
     1441 ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg
     1501 aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt
     1561 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag
     1621 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
     1681 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg
     1741 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa
     1801 taattttgtt taactttaag aaggagatat accatgcatc atcatcatca tcacagcagc
     1861 ggcagagaaa acttgtattt ccagggcgac tccctgtcca tccaccagct cgcagcacag
     1921 ggggagctgg accagctgaa ggagcatttg cggaaaggtg acaacctcgt caacaagcca
     1981 gacgagcgcg gcttcacccc cctcatctgg gcctccgcct ttggagagat tgagaccgtt
     2041 cgcttcctgc tggagtgggg tgccgacccc cacatcctgg caaaagagcg agagagcgcc
     2101 ctgtcgctgg ccagcacagg cggctacaca gacattgtgg ggctgctgct ggagcgtgac
     2161 gtggacatca acatctatga ttggaatgga gggacgccac tgctgtacgc tgtgcgcggg
     2221 aaccacgtga aatgcgttga ggccttgctg gcccgaggcg ctgacctcac caccgaagcc
     2281 gactctggct acaccccgat ggaccttgcc gtggccctgg gataccggaa agtgcaacag
     2341 gtgatcgaga accacatcct caagctcttc cagagcaacc tggtgcccgc tgaccctgag
     2401 tgatgacgaa gcttgcggcc gcactcgagc accaccacca ccaccactga gatccggctg
     2461 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
     2521 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
     2581 ccggattggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2641 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2701 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2761 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     2821 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     2881 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     2941 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3001 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     3061 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3121 atgtatccgc tcatgaatta attcttagaa aaactcatcg agcatcaaat gaaactgcaa
     3181 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
     3241 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
     3301 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
     3361 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc
     3421 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac
     3481 caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa
     3541 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac
     3601 aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat
     3661 cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag
     3721 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac
     3781 gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata
     3841 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc
     3901 atccatgttg gaatttaatc gcggcctaga gcaagacgtt tcccgttgaa tatggctcat
     3961 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg accaaaatcc
     4021 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4081 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4141 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4201 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     4261 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4321 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4381 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4441 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4501 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4561 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4621 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4681 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     4741 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     4801 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     4861 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag
     4921 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     4981 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     5041 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     5101 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     5161 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     5221 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     5281 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
     5341 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
     5401 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
     5461 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
     5521 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
     5581 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
     5641 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
     5701 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc
     5761 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgac
//
