LOCUS       sequence-464165-        6083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Heterologous protein expression of peroxiredoxin-2 (Prx) in 
            Escherichia coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6083)
  TITLE     AI Proteins, Inc. plasmids
REFERENCE   2  (bases 1 to 6083)
  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_1TP9
FEATURES             Location/Qualifiers
     source          1..6083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             421..612
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(606..628)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    714..853
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     769..788
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(868..885)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1039..1627)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1119..1138)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1738..1757)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1749..2564
                     /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             1749..2564
                     /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(1822..1841)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(2656..3111)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2793..2814)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3005..3024
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(3148..3195)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3191..3209
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(3262..3279)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4090..4119)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     RBS             4142..4164
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4179..4203
                     /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(4203..4222)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4204..4222)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4310..4329)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        4535..4612
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4613..5695
                     /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(4632..4651)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tggcggggtt gccttactgg ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc
       61 gactgctgct gcaaaacgtc tgcgacctga gcaacaacat gaatggtctt cggtttccgt
      121 gtttcgtaaa gtctggaaac gcggaagtca gcgccctgca ccattatgtt ccggatctgc
      181 atcgcaggat gctgctggct accctgtgga acacctacat ctgtattaac gaagcgctgg
      241 cattgaccct gagtgatttt tctctggtcc cgccgcatcc ataccgccag ttgtttaccc
      301 tcacaacgtt ccagtaaccg ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc
      361 tctcgtttca tcggtatcat tacccccatg aacagaaatc ccccttacac ggaggcatca
      421 gtgaccaaac aggaaaaaac cgcccttaac atggcccgct ttatcagaag ccagacatta
      481 acgcttctgg agaaactcaa cgagctggac gcggatgaac aggcagacat ctgtgaatcg
      541 cttcacgacc acgctgatga gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt
      601 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
      661 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc
      721 atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc
      781 agattgtact gagagtgcac cattgcggtg tgaaataccg cacagatgcg taaggagaaa
      841 ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
      901 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
      961 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1021 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1081 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1141 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1201 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1261 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1321 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1381 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1441 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     1501 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1561 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1621 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1681 acgttaaggg attttggtca tgaacaataa aactgtctgc ttacataaac agtaatacaa
     1741 ggggtgttat gagccatatt caacgggaaa cgtcttgctc taggccgcga ttaaattcca
     1801 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
     1861 cgacaatcta tcgattgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
     1921 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     1981 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     2041 ccactgcgat ccccgggaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     2101 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     2161 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata
     2221 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     2281 tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     2341 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     2401 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     2461 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     2521 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaagaatta attcatgagc
     2581 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     2641 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     2701 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     2761 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     2821 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     2881 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     2941 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     3001 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     3061 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc
     3121 caatccggat atagttcctc ctttcagcaa aaaacccctc aagacccgtt tagaggcccc
     3181 aaggggttat gctagttatt gctcagcggt ggcagcagcc aactcagctt cctttcgggc
     3241 tttgttagca gccggatctc agtggtggtg gtggtggtgc tcgagtcatt acgaacccag
     3301 gtccttcaaa atatcttccg cagaggacac ggtgaactcg ccgccacctt caatatttgc
     3361 cgccttcacc ttcaagtcgt ccaccagcaa agcaaaacgg cggctacgag tacctaaacc
     3421 tttctcttgg aggtccagtt ccagacccag cgcatgggtg taggtagcgc tgccatccgc
     3481 caggaatttt acgtgtttgt tttccgggta ggattttgcc caggctttca taacaaacgg
     3541 atcgttgacg ctaatacaca ggatctcggt gacgcctttc gacttcagct caccggcctt
     3601 ctcgataaaa cccggcacgt gcttaaggct gcacgtcggg gtaaacgcac ccggaacgcc
     3661 gaacaagatc actttcttgc ctgcaaccag gctatgaaca gaaacctctt gcagctgatc
     3721 ctgttcgtcg aaatacgcca gtttaccatc tggtagaaca tctcctacag ctatgggtgc
     3781 cattccagag ccgccaatct gttcacggtg ggcttcaatg atgtcgttgt cttccatgtc
     3841 caggtcttcc ggcgcttgat ctgcctggat gcgaataccg tcatacagga aacgcaggct
     3901 atccatttcc ttgccctgac gtttggcaaa cgcttccatc aggcgacgca gcggcgtggt
     3961 ctttttgatc ttaaagaaaa tttcagagct gccgtcacta actttcaggt taatatgggt
     4021 ttccggtttc acttccggct tgacttccgg ctttgcttct tggttgactt ccgaatcttg
     4081 caggctaccg tggtgatggt gatggtgatg atggtgatga atcttcgcca tcatatgtat
     4141 atctccttct taaagttaaa caaaattatt tctagagggg aattgttatc cgctcacaat
     4201 tcccctatag tgagtcgtat taatttcgcg ggatcgagat cgatctcgat cctctacgcc
     4261 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
     4321 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc
     4381 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca
     4441 ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg
     4501 caggagtcgc ataagggaga gcgtcgagat cccggacacc atcgaatggc gcaaaacctt
     4561 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc
     4621 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
     4681 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
     4741 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct
     4801 gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat
     4861 taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg
     4921 cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat
     4981 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt
     5041 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca
     5101 tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc
     5161 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa
     5221 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
     5281 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
     5341 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
     5401 cgttggtgcg gacatctcgg tagtgggata cgacgatacc gaagacagct catgttatat
     5461 cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
     5521 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact
     5581 ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     5641 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     5701 acgcaattaa tgtaagttag ctcactcatt aggcaccggg atctcgaccg atgcccttga
     5761 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac
     5821 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca
     5881 ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat
     5941 tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg
     6001 gcgagaagca ggccattatc gccggcatgg cggccccacg ggtgcgcatg atcgtgctcc
     6061 tgtcgttgag gacccggcta ggc
//
