LOCUS       sequence-267726-        5805 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human BP1 fragment of SREBF1 with GB1 solubility tag in vector for 
            in vitro expression, with T7 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5805)
  TITLE     Wagner Lab plasmids
REFERENCE   2  (bases 1 to 5805)
  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: pHIS-GB1-BP1
FEATURES             Location/Qualifiers
     source          1..5805
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     26..45
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        26..44
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    45..69
                     /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             84..106
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             126..143
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             195..356
                     /codon_start=1
                     /product="B1 domain of Streptococcal protein G   "
                     /label=GB1
                     /note="effective as a solubilizing fusion partner (Cheng 
                     and Patel, 2004)"
                     /translation="YKLILNGKTLKGETTTEAVDAATAEKVFKQYANDNGVDGEWTYDD
                     ATKTFTVTE"
     CDS             369..389
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             562..579
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(632..650)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      646..693
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      730..1185
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(817..836)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1027..1048
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1211..1315
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1316..1384,1385..2176)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1534..1553)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2347..2935
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2836..2855
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3089..3106
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3121..3260
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3186..3205)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3346..3368
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3362..3553)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4362..5444)
                     /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     5406..5425
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5445..5522)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5728..5747
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gatcgagatc tcgatcccgc gaaattaata cgactcacta taggggaatt gtgagcggat
       61 aacaattccc ctctagaaat aattttgttt aactttaaga aggagatata catatgtcgt
      121 actaccatca ccatcaccat cacgattacg atatcccaac gaccgaaaac ctgtattttc
      181 tgggcgccat ggagtacaag cttatcctga acggtaaaac cctgaaaggt gaaaccacca
      241 ccgaagctgt tgacgctgct accgcggaaa aagttttcaa acagtacgct aacgacaacg
      301 gtgttgacgg tgaatggacc tacgacgacg ctaccaaaac cttcacggta accgaaatcc
      361 caacgaccga aaacctgtat tttcagggcg aattcgaacc gtgcgatctg gatgcggcgc
      421 tgctgaccga cattgaagat atgctgcaac tgattaacaa tcaggatagc gactaatgag
      481 agctcaacta gtgcggccgc tttcgaatct agagcctgca gtctcgaggc atgcggtacc
      541 aagcttgcgg ccgcactcga gcaccaccac caccaccact gagatccggc tgctaacaaa
      601 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
      661 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggattg
      721 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
      781 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
      841 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
      901 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
      961 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     1021 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     1081 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     1141 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     1201 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     1261 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     1321 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     1381 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     1441 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     1501 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     1561 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     1621 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     1681 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     1741 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     1801 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
     1861 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     1921 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     1981 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     2041 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     2101 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     2161 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     2221 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     2281 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2341 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2401 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2461 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     2521 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2581 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2641 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2701 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     2761 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     2821 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     2881 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     2941 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     3001 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     3061 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     3121 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact
     3181 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     3241 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3301 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3361 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     3421 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     3481 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     3541 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac
     3601 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac
     3661 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca
     3721 ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc
     3781 agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca
     3841 gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt
     3901 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa
     3961 ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg
     4021 gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag
     4081 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca
     4141 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct
     4201 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
     4261 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg
     4321 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
     4381 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4441 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg
     4501 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc
     4561 aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta
     4621 tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg
     4681 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc
     4741 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt
     4801 tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga
     4861 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg
     4921 accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg
     4981 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca
     5041 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg
     5101 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc
     5161 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc
     5221 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt
     5281 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc
     5341 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag
     5401 acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat
     5461 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg
     5521 tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag
     5581 gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa
     5641 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
     5701 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
     5761 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagag
//
