LOCUS       sequence-137195-        6391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression for structure determination; may not be full 
            ORF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6391)
  TITLE     SGC Oxford
REFERENCE   2  (bases 1 to 6391)
  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: TPH2
FEATURES             Location/Qualifiers
     source          1..6391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     CDS             91..108
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             133..153
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1251..1268
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1322..1340)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1336..1383
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1420..1875
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1507..1526)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1717..1738
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1968..2783)
                     /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(1968..2783)
                     /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     2691..2710
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2775..2794
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2905..3493
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3394..3413
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3647..3664
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3679..3821
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3747..3766)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3907..3929
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3923..4114)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4923..6005)
                     /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     5967..5986
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6006..6083)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6289..6308
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt tagaaggaga tatacatatg caccatcatc atcatcattc ttctggtgta
      121 gatctgggta ccgagaacct gtacttccaa tccatgctag aggatgtgcc ctggttccct
      181 cggaagatct ctgagttaga caaatgctct cacagagttc tcatgtatgg ttctgagctt
      241 gatgctgacc acccaggatt taaggacaat gtctatcgac agagaagaaa gtattttgtg
      301 gatgtggcca tgggttataa atatggtcag cccattccca gggtggagta tactgaagaa
      361 gaaactaaaa cttggggtgt tgtattccgg gagctctcca aactctatcc cactcatgct
      421 tgccgagagt atttgaaaaa cttccctctg ctgactaaat actgtggcta cagagaggac
      481 aatgtgcctc aactcgaaga tgtctccatg tttctgaaag aaaggtctgg cttcacggtg
      541 aggccggtgg ctggatacct gagcccacga gactttctgg caggactggc ctacagagtg
      601 ttccactgta cccagtacat ccggcatggc tcagatcccc tctacacccc ggaaccagac
      661 acatgccatg aactcttggg acatgttcca ctacttgcgg atcctaagtt tgctcagttt
      721 tcacaagaaa taggtctggc gtctctggga gcatcagatg aagatgttca gaaactagcc
      781 acgtgctatt tcttcacaat cgagtttggc ctttgcaagc aagaagggca actgcgggct
      841 tatggagcag gactcctttc ctccattgga gaattaaagc acgccctttc tgacaaggca
      901 tgtgtgaaag cctttgaccc aaagacaact tgcttacagg aatgccttat caccaccttc
      961 caggaagcct actttgtttc agaaagtttt gaagaagcca aagaaaagat gagggacttt
     1021 gcaaagtcaa ttacccgtcc cttctcagta tacttcaatc cctacacaca gagtattgaa
     1081 attctgaaag acaccagaag tattgaaaat gtggtgcagg accttcgcag cgacttgaat
     1141 acagtgtgtg atgctttaaa caaaatgaac caatatctgg ggatttgaca gtaaaggtgg
     1201 atacggatcc gaattcgagc tccgtcgaca agcttgcggc cgcactcgag caccaccacc
     1261 accaccactg agatcccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     1321 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     1381 ttgctgaaag gaggaactat atccggattg gcgaatggga cgcgccctgt agcggcgcat
     1441 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     1501 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     1561 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     1621 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     1681 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     1741 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     1801 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     1861 taacgtttac aatttcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     1921 tatttttcta aatacattca aatatgtatc cgctcatgaa ttaattctta gaaaaactca
     1981 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     2041 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     2101 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     2161 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     2221 aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     2281 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga
     2341 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     2401 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     2461 tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     2521 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     2581 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     2641 tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
     2701 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac
     2761 gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt
     2821 tttattgttc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     2881 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2941 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3001 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     3061 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3121 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3181 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3241 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3301 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3361 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3421 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3481 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3541 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3601 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3661 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     3721 acaccgcata tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     3781 tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac
     3841 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     3901 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
     3961 agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat
     4021 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg
     4081 ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc
     4141 tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg
     4201 atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc
     4261 ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag
     4321 gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg
     4381 gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg
     4441 ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg
     4501 attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga
     4561 gcacgatcat gcgcacccgt ggggccgcca tgccggcgat aatggcctgc ttctcgccga
     4621 aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata
     4681 ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga
     4741 cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg
     4801 cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca
     4861 agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc
     4921 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4981 aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt
     5041 gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg
     5101 tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata
     5161 taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc
     5221 agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc
     5281 atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg
     5341 gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta
     5401 tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg
     5461 atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg
     5521 gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca
     5581 ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc
     5641 agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg
     5701 cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc
     5761 gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc
     5821 aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc
     5881 atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg
     5941 cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt
     6001 ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag
     6061 gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat
     6121 taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc
     6181 atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
     6241 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
     6301 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
     6361 gtagaggatc gagatctcga tcccgcgaaa t
//
