LOCUS       sequence-464157-        5903 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Heterologous protein expression of aprataxin in Escherichia coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5903)
  TITLE     AI Proteins, Inc. plasmids
REFERENCE   2  (bases 1 to 5903)
  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: pAIP4_3KT9
FEATURES             Location/Qualifiers
     source          1..5903
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             322..513
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(507..529)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    615..754
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     670..689
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(769..786)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(940..1528)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1020..1039)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1639..1658)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1650..2465
                     /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             1650..2465
                     /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     complement(1723..1742)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(2557..3012)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2694..2715)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2906..2925
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(3049..3096)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3092..3110
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(3163..3180)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(3811..3840)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     RBS             3863..3885
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    3900..3924
                     /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)."
     primer_bind     complement(3924..3943)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3925..3943)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4031..4050)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        4256..4333
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4334..5416
                     /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     complement(4353..4372)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc
       61 accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca
      121 tctgtattaa cgaagcgctg gcattgaccc tgagtgattt ttctctggtc ccgccgcatc
      181 cataccgcca gttgtttacc ctcacaacgt tccagtaacc gggcatgttc atcatcagta
      241 acccgtatcg tgagcatcct ctctcgtttc atcggtatca ttacccccat gaacagaaat
      301 cccccttaca cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc
      361 tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga cgcggatgaa
      421 caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg cagctgcctc
      481 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
      541 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
      601 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc
      661 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccattgcggt gtgaaatacc
      721 gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga
      781 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
      841 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
      901 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
      961 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1021 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1081 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1141 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1201 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1261 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1321 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1381 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1441 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1501 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     1561 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgaacaata aaactgtctg
     1621 cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcttgct
     1681 ctaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
     1741 ataatgtcgg gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag
     1801 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
     1861 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
     1921 ctgatgatgc atggttactc accactgcga tccccgggaa aacagcattc caggtattag
     1981 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
     2041 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc
     2101 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
     2161 atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg
     2221 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     2281 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     2341 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     2401 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
     2461 tctaagaatt aattcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     2521 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt
     2581 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     2641 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     2701 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     2761 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     2821 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     2881 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     2941 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     3001 tacagggcgc gtcccattcg ccaatccgga tatagttcct cctttcagca aaaaacccct
     3061 caagacccgt ttagaggccc caaggggtta tgctagttat tgctcagcgg tggcagcagc
     3121 caactcagct tcctttcggg ctttgttagc agccggatct cagtggtggt ggtggtggtg
     3181 ctcgagtcat tacgaacctt cctcgaactc cacgatgtac gggtacagct cgttgaccat
     3241 gtgcagtacc tgacccggct gcaacttaac ttcttggtcc ttgccgatca caacgctatc
     3301 aatggacgtc gggttcacgc ccacttgctt caccttaaca taacctttat tacattccgc
     3361 tttcagctgg acttgctggc ggctgcactt tttatcggtg attttggttt ccggaccacg
     3421 gccaataaca acagcctcca agtgtggcag acgaatacgc tgatggcgag agtcttgtct
     3481 aactagccaa cataccctca tcattccaga gccgccaatc tgttcacggt gggcttcaat
     3541 gatgtcgttg tcttccatgt ccaggtcttc cggcgcttga tctgcctgga tgcgaatacc
     3601 gtcatacagg aaacgcaggc tatccatttc cttgccctga cgtttggcaa acgcttccat
     3661 caggcgacgc agcggcgtgg tctttttgat cttaaagaaa atttcagagc tgccgtcact
     3721 aactttcagg ttaatatggg tttccggttt cacttccggc ttgacttccg gctttgcttc
     3781 ttggttgact tccgaatctt gcaggctacc gtggtgatgg tgatggtgat gatggtgatg
     3841 aatcttcgcc atcatatgta tatctccttc ttaaagttaa acaaaattat ttctagaggg
     3901 gaattgttat ccgctcacaa ttcccctata gtgagtcgta ttaatttcgc gggatcgaga
     3961 tcgatctcga tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg
     4021 gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg
     4081 ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg
     4141 ggcgccatct ccttgcatgc accattcctt gcggcggcgg tgctcaacgg cctcaaccta
     4201 ctactgggct gcttcctaat gcaggagtcg cataagggag agcgtcgaga tcccggacac
     4261 catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt
     4321 cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc
     4381 ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga
     4441 aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact
     4501 ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc
     4561 gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt
     4621 gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc
     4681 gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt
     4741 ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat
     4801 caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc
     4861 attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct
     4921 gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg
     4981 ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg
     5041 catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc
     5101 cattaccgag tccgggctgc gcgttggtgc ggacatctcg gtagtgggat acgacgatac
     5161 cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct
     5221 ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa
     5281 tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac
     5341 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     5401 ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcaccgg
     5461 gatctcgacc gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg
     5521 gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg
     5581 tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga
     5641 tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg
     5701 gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccccac
     5761 gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg ggttgcctta
     5821 ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa
     5881 cgtctgcgac ctgagcaaca aca
//
