LOCUS       sequence-464191-        6086 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Heterologous protein expression of peptidyl-prolyl cis-trans 
            isomerase A (PPIase A) in Escherichia coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6086)
  TITLE     AI Proteins, Inc. plasmids
REFERENCE   2  (bases 1 to 6086)
  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_5F66
FEATURES             Location/Qualifiers
     source          1..6086
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             260..451
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(445..467)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    553..692
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     608..627
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(707..724)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(878..1466)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(958..977)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1577..1596)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1588..2403
                     /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             1588..2403
                     /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(1661..1680)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(2495..2950)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2632..2653)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2844..2863
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(2987..3034)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3030..3048
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(3101..3118)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(3932..3961)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     RBS             3984..4006
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4021..4045
                     /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(4045..4064)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4046..4064)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4152..4171)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        4377..4454
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4455..5537
                     /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(4474..4493)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 cattatgttc cggatctgca tcgcaggatg ctgctggcta ccctgtggaa cacctacatc
       61 tgtattaacg aagcgctggc attgaccctg agtgattttt ctctggtccc gccgcatcca
      121 taccgccagt tgtttaccct cacaacgttc cagtaaccgg gcatgttcat catcagtaac
      181 ccgtatcgtg agcatcctct ctcgtttcat cggtatcatt acccccatga acagaaatcc
      241 cccttacacg gaggcatcag tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt
      301 tatcagaagc cagacattaa cgcttctgga gaaactcaac gagctggacg cggatgaaca
      361 ggcagacatc tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca gctgcctcgc
      421 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
      481 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
      541 cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt
      601 aactatgcgg catcagagca gattgtactg agagtgcacc attgcggtgt gaaataccgc
      661 acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact
      721 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
      781 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
      841 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
      901 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
      961 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     1021 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     1081 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     1141 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     1201 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     1261 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     1321 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     1381 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     1441 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     1501 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gaacaataaa actgtctgct
     1561 tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac gtcttgctct
     1621 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     1681 aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga tgcgccagag
     1741 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     1801 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     1861 gatgatgcat ggttactcac cactgcgatc cccgggaaaa cagcattcca ggtattagaa
     1921 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     1981 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     2041 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     2101 ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat
     2161 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     2221 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     2281 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     2341 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     2401 taagaattaa ttcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     2461 ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta
     2521 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     2581 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     2641 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     2701 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     2761 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     2821 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     2881 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     2941 cagggcgcgt cccattcgcc aatccggata tagttcctcc tttcagcaaa aaacccctca
     3001 agacccgttt agaggcccca aggggttatg ctagttattg ctcagcggtg gcagcagcca
     3061 actcagcttc ctttcgggct ttgttagcag ccggatctca gtggtggtgg tggtggtgct
     3121 cgagtcatta cgaaccttcc agctgaccgc agtccgcaat ggtgatcttt tttgaagtct
     3181 tgccattacg gctaccaaaa cgttccatag cttccacgat gttcatacct tctttgacct
     3241 taccaaaaac cacgtgcttg ccatccagcc attccgtttt cgcggtgcag ataaaaaatt
     3301 gcgagccgtt ggtatttgga cccgcatttg ccatgctcag aatacccgga ccggtgtgtt
     3361 tgaggatgaa gttctcatcc tcgaacttct cgccgtagat gctcttgcca ccggtgccgt
     3421 tatggcgggt gaaatcacca ccctgacaca tgaaacccgg aataatacga tggaagcagc
     3481 tgcctttata gccgaagccc ttctcacccg tagacaacgc gcggaagttt tctgcggttt
     3541 tcggaacttt gtcggcaaac agctcaaagg acacacgacc taacggctcg ccgtctacag
     3601 ctatatcgaa aaaaactgtg ggatttccag agccgccaat ctgttcacgg tgggcttcaa
     3661 tgatgtcgtt gtcttccatg tccaggtctt ccggcgcttg atctgcctgg atgcgaatac
     3721 cgtcatacag gaaacgcagg ctatccattt ccttgccctg acgtttggca aacgcttcca
     3781 tcaggcgacg cagcggcgtg gtctttttga tcttaaagaa aatttcagag ctgccgtcac
     3841 taactttcag gttaatatgg gtttccggtt tcacttccgg cttgacttcc ggctttgctt
     3901 cttggttgac ttccgaatct tgcaggctac cgtggtgatg gtgatggtga tgatggtgat
     3961 gaatcttcgc catcatatgt atatctcctt cttaaagtta aacaaaatta tttctagagg
     4021 ggaattgtta tccgctcaca attcccctat agtgagtcgt attaatttcg cgggatcgag
     4081 atcgatctcg atcctctacg ccggacgcat cgtggccggc atcaccggcg ccacaggtgc
     4141 ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg
     4201 gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt
     4261 gggcgccatc tccttgcatg caccattcct tgcggcggcg gtgctcaacg gcctcaacct
     4321 actactgggc tgcttcctaa tgcaggagtc gcataaggga gagcgtcgag atcccggaca
     4381 ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat
     4441 tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct
     4501 cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg
     4561 aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac
     4621 tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc
     4681 cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg
     4741 tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg
     4801 cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg
     4861 tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca
     4921 tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg
     4981 cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc
     5041 tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac
     5101 gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg
     5161 gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg
     5221 ccattaccga gtccgggctg cgcgttggtg cggacatctc ggtagtggga tacgacgata
     5281 ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc
     5341 tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca
     5401 atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa
     5461 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     5521 tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt agctcactca ttaggcaccg
     5581 ggatctcgac cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
     5641 ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
     5701 gtgccggcag cgctctgggt cattttcggc gaggaccgct ttcgctggag cgcgacgatg
     5761 atcggcctgt cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc cttcgtcact
     5821 ggtcccgcca ccaaacgttt cggcgagaag caggccatta tcgccggcat ggcggcccca
     5881 cgggtgcgca tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt
     5941 actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa
     6001 acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg
     6061 gaaacgcgga agtcagcgcc ctgcac
//
