LOCUS       sequence-137211-        6577 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 6577)
  TITLE     SGC Oxford
REFERENCE   2  (bases 1 to 6577)
  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: SEC14L4
FEATURES             Location/Qualifiers
     source          1..6577
                     /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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             92..109
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             134..154
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1438..1455
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1508..1526)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1522..1569
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1606..2061
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1693..1712)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1903..1924
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2154..2969)
                     /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(2154..2969)
                     /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     2877..2896
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2961..2980
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3091..3679
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3580..3599
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3833..3850
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3865..4007
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3933..3952)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4093..4115
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4109..4300)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5109..6191)
                     /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     6153..6172
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6192..6269)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6475..6494
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gcaccatcat catcatcatt cttctggtgt
      121 agatctgggt accgagaacc tgtacttcca atccatgtcg tcccgtgtgg gtgacctgtc
      181 gccgcaacaa caagaagccc tggcccgctt ccgtgaaaac ctgcaagacc tgctgccgat
      241 tctgccgaac gcggatgact attttctgct gcgttggctg cgtgcccgca atttcgatct
      301 gcagaaaagt gaagacatgc tgcgtcgtca catggaattt cgcaaacagc aggatctgga
      361 taacattgtc acctggcagc cgccggaagt gatccaactg tatgattcag gcggtctgtg
      421 cggctatgac tacgaaggtt gtccggtgta cttcaatatt atcggcagcc tggacccgaa
      481 aggtctgctg ctgagtgctt ccaagcagga catgattcgt aaacgcatca aggtgtgcga
      541 actgctgctg catgaatgtg aactgcagac ccaaaaactg ggccgtaaga ttgaaatggc
      601 gctgatggtt tttgatatgg aaggtctgag tctgaaacac ctgtggaagc cggcagtgga
      661 agtttatcag caatttttct ccatcctgga agctaactac ccggaaacgc tgaaaaatct
      721 gattgttatc cgcgcgccga agctgtttcc ggtggccttc aacctggtta aatcattcat
      781 gtcggaagaa acccgtcgca aaattgtgat cctgggcgat aattggaaac aggaactgac
      841 gaagtttatt agcccggacc aactgccggt tgaatttggc ggcaccatga ccgatccgga
      901 cggtaacccg aaatgcctga ccaagatcaa ttatggcggt gaagtgccga aaagctatta
      961 cctgtgtgaa caggttcgtc tgcaatacga acacacgcgt tctgttggcc gcggtagctc
     1021 tctgcaggtc gaaaacgaaa ttctgtttcc gggttgcgtt ctgcgttggc agttcgcaag
     1081 cgatggcggt gacatcggct ttggtgtctt cctgaaaacc aagatgggtg aacagcaatc
     1141 agcgcgtgaa atgacggaag tcctgccgtc gcagcgctat aacgcccaca tggtgccgga
     1201 agatggctct ctgacctgtc tgcaggcagg tgtctatgtg ctgcgttttg ataataccta
     1261 cagtcgcatg cacgcgaaaa agctgtccta cacggttgaa gtcctgctgc cggataaagc
     1321 aagcgaagaa accctgcagt ctctgaaagc tatgcgtccg agcccgacgc aatgacagta
     1381 aaggtggata cggatccgaa ttcgagctcc gtcgacaagc ttgcggccgc actcgagcac
     1441 caccaccacc accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct
     1501 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     1561 ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg
     1621 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     1681 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     1741 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     1801 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     1861 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     1921 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     1981 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     2041 aaatattaac gtttacaatt tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2101 tttgtttatt tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa
     2161 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
     2221 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     2281 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     2341 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     2401 ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta
     2461 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
     2521 gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
     2581 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     2641 aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga
     2701 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
     2761 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     2821 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
     2881 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag
     2941 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
     3001 gacagtttta ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3061 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3121 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3181 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     3241 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3301 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3361 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3421 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3481 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3541 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3601 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3661 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3721 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3781 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     3841 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     3901 tatttcacac cgcatatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     3961 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
     4021 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     4081 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     4141 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct
     4201 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
     4261 agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg
     4321 gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg
     4381 ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat
     4441 ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag
     4501 atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg
     4561 tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc
     4621 atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta
     4681 tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg
     4741 acaggagcac gatcatgcgc acccgtgggg ccgccatgcc ggcgataatg gcctgcttct
     4801 cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc
     4861 cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga
     4921 aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca
     4981 taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg
     5041 ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg
     5101 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5161 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc
     5221 accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc
     5281 aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc
     5341 gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca
     5401 acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca
     5461 accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg
     5521 gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga
     5581 tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac
     5641 agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct
     5701 tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc
     5761 ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta
     5821 atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg
     5881 acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat
     5941 ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca
     6001 atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc
     6061 tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc
     6121 accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt
     6181 actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg
     6241 cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc
     6301 ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa
     6361 tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc
     6421 cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
     6481 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
     6541 tccggcgtag aggatcgaga tctcgatccc gcgaaat
//
