LOCUS       sequence-319830-        5566 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Baculovirus expression for structure determination.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5566)
  TITLE     SGC Toronto 2019-Q1 to 2020-Q2
REFERENCE   2  (bases 1 to 5566)
  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: 6OGJ
FEATURES             Location/Qualifiers
     source          1..5566
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             320..337
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             387..404
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(457..475)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      471..518
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      555..1010
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(642..661)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     852..873
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1102..1917)
                     /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(1102..1917)
                     /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     1825..1844
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1909..1928
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2039..2627
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2528..2547
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2781..2798
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2813..2952
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2878..2897)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3038..3060
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3054..3245)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4054..5136)
                     /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     5098..5117
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5137..5214)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5420..5439
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5523..5542
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5523..5541
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5542..5566
                     /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)."
ORIGIN
        1 cctctagaaa taattttgtt taaccttaag aaggagatat actatggctt ctgcctcccc
       61 caaacagcgg cgctccatca tccgtgaccg gggacccatg tatgatgacc ccaccctgcc
      121 tgaaggctgg acacggaagc ttaagcaaag gaaatctggc cgctctgctg ggaagtatga
      181 tgtgtatttg atcaatcccc agggaaaagc ctttcgctct aaagtggagt tgattgcgta
      241 cttcgaaaag gtaggcgaca catccctgga ccctaatgat tttgacttca cggtaactgg
      301 gagagggagc ccctccgcgc accatcatca ccaccattga ggatccgaat tcgagctccg
      361 tcgacaagct tgcggccgca ctcgagcacc accaccacca ccactgagat ccggctgcta
      421 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
      481 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg
      541 gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
      601 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
      661 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
      721 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
      781 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
      841 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
      901 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
      961 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
     1021 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     1081 tatccgctca tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt
     1141 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
     1201 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
     1261 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
     1321 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt
     1381 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
     1441 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg
     1501 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
     1561 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc
     1621 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     1681 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     1741 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat
     1801 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     1861 catgttggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcataac
     1921 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgacc aaaatccctt
     1981 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2041 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2101 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2161 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     2221 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2281 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2341 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2401 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     2461 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     2521 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     2581 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     2641 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     2701 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     2761 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     2821 ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac
     2881 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
     2941 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     3001 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     3061 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
     3121 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
     3181 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
     3241 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
     3301 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
     3361 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
     3421 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
     3481 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac
     3541 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag
     3601 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc
     3661 cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg tggggccgcc
     3721 atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag
     3781 gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg
     3841 ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg
     3901 agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac
     3961 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa
     4021 tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4081 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4141 gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac
     4201 cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa
     4261 atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta
     4321 tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc
     4381 gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag
     4441 catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat
     4501 cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga
     4561 gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg
     4621 ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg
     4681 gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc
     4741 atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt
     4801 gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct
     4861 ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag
     4921 ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc
     4981 cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt
     5041 cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc
     5101 atactctgcg acatcgtata acgttactgg tttcacattc accaccctga attgactctc
     5161 ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat
     5221 ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc
     5281 cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc
     5341 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
     5401 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
     5461 cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa
     5521 attaatacga ctcactatag gggaattgtg agcggataac aattcc
//
