LOCUS       sequence-361434-        6445 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 6445)
  TITLE     SGC Structures-2021-Q3 and Q4
REFERENCE   2  (bases 1 to 6445)
  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: PF10_0086:MAC044-A01:C220746
FEATURES             Location/Qualifiers
     source          1..6445
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     48..67
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     151..170
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        151..169
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    170..194
                     /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             209..231
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             241..258
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             271..291
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1096..1114)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1110..1157
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1332..1337,1338..1354,1355..1360))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1387..1405)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1473..1577
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1578..1646,1647..2438)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1796..1815)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2609..3197
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3098..3117
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3351..3368
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3383..3522
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3448..3467)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3608..3630
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3624..3815)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5127..6209)
                     /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     6171..6190
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6210..6287)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg
       61 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg
      121 tagaggatcg agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag
      181 cggataacaa ttcccctcta gaaataattt tgtttaactt taagaaggag atataccatg
      241 catcatcatc atcatcacag cagcggcaga gaaaacttgt atttccaggg catgaatgaa
      301 aatttagaaa atttttcaac aattgatttg ttgaatgaat taaagaggag gtatgcatgt
      361 ttgagtaaac cagatggtag atatattttt ttaggtgccc caggatctgg aaaaggaaca
      421 caatccttga accttaagaa gtctcattgt tactgtcatt tatcaacagg tgatttgtta
      481 agagaagcag ctgagaaaaa aacggaactt ggacttaaaa ttaagaacat cattaatgaa
      541 gggaaattag ttgatgatca aatggtattg tcactagtcg atgagaaatt aaaaacacct
      601 caatgtaaaa aaggatttat tcttgatgga tatccaagaa atgtaaaaca agctgaagat
      661 ttaaataagc tactacaaaa aaatcaaaca aaattagacg gtgtgtttta ttttaatgta
      721 cctgatgaag ttttagttaa tagaattagt ggtagattaa ttcataaacc ttccggaaga
      781 atttatcata aaatctttaa tcctccaaaa gttcccttta gagatgatgt aacaaacgaa
      841 ccattaatac aaagagaaga tgataatgaa gatgttttaa aaaaaagact aacggttttt
      901 aaaagcgaaa catcaccttt aataagttat tataaaaata aaaatttact tattaattta
      961 gacgcaacac aaccagcaaa tgatttggaa aaaaaaatct ctcaacatat agacggttga
     1021 tgacgaagct tgcggccgca ctcgaggatc cggctgctaa caaagcccga aaggaagctg
     1081 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
     1141 tcttgagggg ttttttgctg aaaggaggaa ctatatccgg atatcccgca agaggcccgg
     1201 cagtaccggc ataaccaagc ctatgcctac agcatccagg gtgacggtgc cgaggatgac
     1261 gatgagcgca ttgttagatt tcatacacgg tgcctgactg cgttagcaat ttaactgtga
     1321 taaactaccg cattaaagct tatcgatgat aagctgtcaa acatgagaat tcttgaagac
     1381 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     1441 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1501 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     1561 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     1621 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     1681 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     1741 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     1801 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     1861 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     1921 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     1981 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2041 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2101 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2161 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     2221 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     2281 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     2341 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     2401 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     2461 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     2521 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     2581 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     2641 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     2701 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     2761 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     2821 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2881 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2941 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3001 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3061 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3121 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3181 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3241 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3301 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     3361 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     3421 tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     3481 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     3541 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     3601 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     3661 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     3721 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     3781 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     3841 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     3901 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     3961 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta
     4021 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag
     4081 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt
     4141 gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt
     4201 gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg
     4261 agcacgatca tgcgcacccg tggccaggac ccaacgctgc ccgagatgcg ccgcgtgcgg
     4321 ctgctggaga tggcggacgc gatggatatg ttctgccaag ggttggtttg cgcattcaca
     4381 gttctccgca agaattgatt ggctccaatt cttggagtgg tgaatccgtt agcgaggtgc
     4441 cgccggcttc cattcaggtc gaggtggccc ggctccatgc accgcgacgc aacgcgggga
     4501 ggcagacaag gtatagggcg gcgcctacaa tccatgccaa cccgttccat gtgctcgccg
     4561 aggcggcata aatcgccgtg acgatcagcg gtccaatgat cgaagttagg ctggtaagag
     4621 ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc tacctgcctg gacagcatgg
     4681 cctgcaacgc gggcatcccg atgccgccgg aagcgagaag aatcataatg gggaaggcca
     4741 tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag cgcgtcggcc gccatgccgg
     4801 cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag
     4861 cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc
     4921 gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca
     4981 tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg
     5041 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga
     5101 gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5161 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc
     5221 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
     5281 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt
     5341 ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact
     5401 accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc
     5461 gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc
     5521 atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga
     5581 atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa
     5641 cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg
     5701 cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag
     5761 acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg
     5821 tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc
     5881 gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc
     5941 agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga
     6001 ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg
     6061 ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa
     6121 acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct
     6181 gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg
     6241 cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg
     6301 ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag
     6361 caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac
     6421 ggggcctgcc accataccca cgccg
//
