LOCUS       sequence-137200-        6312 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 6312)
  TITLE     SGC Oxford
REFERENCE   2  (bases 1 to 6312)
  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: MAGEA3
FEATURES             Location/Qualifiers
     source          1..6312
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             44..61
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             71..397
                     /codon_start=1
                     /gene="trxA"
                     /product="E. coli thioredoxin"
                     /label=TrxA
                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
                     IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
                     DANLA"
     CDS             404..424
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1126..1143
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1196..1214)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1210..1257
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1294..1749
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1381..1400)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1591..1612
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1842..2657)
                     /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(1842..2657)
                     /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     2565..2584
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2649..2668
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2779..3367
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3268..3287
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3521..3538
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3553..3695
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3621..3640)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3781..3803
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3797..3988)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4797..5879)
                     /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     5841..5860
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5880..5957)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6163..6182
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6266..6285
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6266..6284
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6285..6309
                     /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)."
ORIGIN
        1 ctagaaataa ttttgtttaa ctttagaagg agatatacat atgcaccatc atcatcatca
       61 ttcttctggt atgagcgata aaattattca cctgactgac gacagttttg acacggatgt
      121 actcaaagcg gacggggcga tcctcgtcga tttctgggca gagtggtgcg gtccgtgcaa
      181 aatgatcgcc ccgattctgg atgaaatcgc tgacgaatat cagggcaaac tgaccgttgc
      241 aaaactgaac atcgatcaaa accctggcac tgcgccgaaa tatggcatcc gtggtatccc
      301 gactctgctg ctgttcaaaa acggtgaagt ggcggcaacc aaagtgggcg cactgtctaa
      361 aggtcagttg aaagagttcc tcgacgctaa cctggccggt accgagaact tgtacttcca
      421 atccatggag ttccaagcag cactcagtag gaaggtggcc gagttggttc attttctgct
      481 cctcaagtat cgagccaggg agccggtcac aaaggcagaa atgctgggga gtgtcgtcgg
      541 aaattggcag tatttctttc ctgtgatctt cagcaaagct tccagttcct tgcagctggt
      601 ctttggcatc gagctgatgg aagtggaccc catcggccac ttgtacatct ttgccacctg
      661 cctgggcctc tcctacgatg gcctgctggg tgacaatcag atcatgccca aggcaggcct
      721 cctgataatc gtcctggcca taatcgcaag agagggcgac tgtgcccctg aggagaaaat
      781 ctgggaggag ctgagtgtgt tagaggtgtt tgaggggagg gaagacagta tcttggggga
      841 tcccaagaag ctgctcaccc aacatttcgt gcaggaaaac tacctggagt accggcaggt
      901 ccccggcagt gatcctgcat gttatgaatt cctgtggggt ccaagggccc tcgttgaaac
      961 cagctatgtg aaagtcctgc accatatggt aaagatcagt ggaggacctc acatttccta
     1021 cccacccctg catgagtggg ttttgagaga gggggaagag tgacagtaaa ggtggatacg
     1081 gatccgaatt cgagctccgt cgacaagctt gcggccgcac tcgagcacca ccaccaccac
     1141 cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     1201 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
     1261 aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     1321 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     1381 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     1441 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     1501 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     1561 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     1621 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     1681 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     1741 ttacaatttc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     1801 tctaaataca ttcaaatatg tatccgctca tgaattaatt cttagaaaaa ctcatcgagc
     1861 atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc
     1921 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg
     1981 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca
     2041 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc
     2101 aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca
     2161 aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat
     2221 acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac
     2281 actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat
     2341 gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa
     2401 tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct
     2461 gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc
     2521 ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta
     2581 tacccatata aatcagcatc catgttggaa tttaatcgcg gcctagagca agacgtttcc
     2641 cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt
     2701 gttcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     2761 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2821 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2881 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     2941 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3001 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3061 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3121 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3181 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3241 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3301 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3361 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3421 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     3481 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     3541 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     3601 catatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
     3661 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
     3721 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     3781 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
     3841 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
     3901 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
     3961 taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca
     4021 tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg
     4081 aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg
     4141 accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc
     4201 cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg
     4261 acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc
     4321 aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat
     4381 tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga
     4441 tcatgcgcac ccgtggggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt
     4501 tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa
     4561 gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga
     4621 gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga
     4681 cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca
     4741 tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac
     4801 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4861 cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg
     4921 ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg
     4981 ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat
     5041 gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg
     5101 gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca
     5161 gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc
     5221 cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag
     5281 ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc
     5341 tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa
     5401 ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg
     5461 caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca
     5521 ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt
     5581 tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg
     5641 acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac
     5701 tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc
     5761 gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa
     5821 acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca
     5881 ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg
     5941 cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa
     6001 gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa
     6061 ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca
     6121 agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata
     6181 ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag
     6241 gatcgagatc tcgatcccgc gaaattaata cgactcacta taggggaatt gtgagcggat
     6301 aacaattccc ct
//
