LOCUS       sequence-361416-        6287 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 6287)
  TITLE     SGC Structures-2021-Q3 and Q4
REFERENCE   2  (bases 1 to 6287)
  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: C22orf4:HPC057-F11:C35657
FEATURES             Location/Qualifiers
     source          1..6287
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     45..64
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     148..167
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        148..166
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    167..191
                     /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             206..228
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             238..255
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             268..288
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1299..1316
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1369..1387)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1383..1430
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1467..1922
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1554..1573)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1764..1785
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2014..2829)
                     /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(2014..2829)
                     /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     2737..2756
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2821..2840
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2951..3539
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3440..3459
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3693..3710
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3725..3864
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3790..3809)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3950..3972
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3966..4157)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4966..6048)
                     /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     6010..6029
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6049..6126)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
       61 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
      121 aggatcgaga tctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg
      181 ataacaattc ccctctagaa ataattttgt ttaactttaa gaaggagata taccatgcat
      241 catcatcatc atcacagcag cggcagagaa aacttgtatt tccagggcag cgaaagagag
      301 gcctcccggc tcgacaagtt caagcagctg cttgccggcc ccaacacgga ccttgaggaa
      361 ttacggaggt tgagctggtc cggaatccct aagccagtgc gtccaatgac gtggaagctc
      421 ctctcaggtt accttcccgc caatgtagac cggagaccag ccactctcca gagaaaacaa
      481 aaagaatatt ttgcatttat tgagcactat tacgattcta ggaacgacga agttcaccag
      541 gacacataca ggcagatcca catagacatc cctcgcatga gccctgaagc gttgatcctg
      601 cagcccaagg tgacggagat ttttgaaagg atcttgttca tatgggcgat ccgccaccca
      661 gccagtggat acgttcaggg tataaatgat ctcgtcactc ctttctttgt ggtcttcatt
      721 tgtgaataca tagaggcaga ggaggtggac acggtggacg tctccggcgt gcccgcagag
      781 gtgctgtgca acatcgaggc cgacacctac tggtgcatga gcaagctgct ggatggcatt
      841 caggacaact acacctttgc ccaacctggg attcaaatga aagtgaaaat gttagaagaa
      901 ctcgtgagcc ggattgatga gcaagtgcac cggcacctgg accaacacga agtgagatac
      961 ctgcagtttg ccttccgctg gatgaacaac ctgctgatga gggaggtgcc cctgcgttgt
     1021 accatccgcc tgtgggacac ctaccagtct gaaccggacg gcttttctca tttccacttg
     1081 tacgtgtgcg ctgcttttct cgtgagatgg aggaaggaaa tactagaaga aaaagatttt
     1141 caagagctgc tgctcttcct ccagaacctg cccacagccc actgggatga tgaggacatc
     1201 agcctgttgc tggccgaggc ctaccgcctc aagtttgctt ttgccgacgc ccccaatcac
     1261 tacaagaaat gatgacgaag cttgcggccg cactcgagca ccaccaccac caccactgag
     1321 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     1381 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     1441 gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
     1501 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     1561 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     1621 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     1681 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     1741 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     1801 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     1861 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     1921 ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     1981 cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga gcatcaaatg
     2041 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
     2101 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     2161 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     2221 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt
     2281 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     2341 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc
     2401 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     2461 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     2521 cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     2581 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     2641 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     2701 caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     2761 taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt cccgttgaat
     2821 atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga
     2881 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     2941 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3001 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3061 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     3121 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3181 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3241 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3301 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3361 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3421 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3481 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3541 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3601 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     3661 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     3721 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg
     3781 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     3841 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     3901 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     3961 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
     4021 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg
     4081 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
     4141 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
     4201 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
     4261 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
     4321 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
     4381 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
     4441 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
     4501 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca
     4561 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc
     4621 cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga
     4681 ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg
     4741 atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc
     4801 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg
     4861 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
     4921 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
     4981 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5041 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     5101 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     5161 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     5221 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
     5281 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     5341 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     5401 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     5461 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     5521 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     5581 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     5641 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     5701 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     5761 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     5821 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     5881 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     5941 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     6001 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     6061 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     6121 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta
     6181 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
     6241 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaa
//
