LOCUS       sequence-464199-        6128 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Heterologous protein expression of peptidyl-prolyl cis-trans 
            isomerase in Escherichia coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6128)
  TITLE     AI Proteins, Inc. plasmids
REFERENCE   2  (bases 1 to 6128)
  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_1YW5
FEATURES             Location/Qualifiers
     source          1..6128
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(77..98)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     289..308
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(432..479)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     475..493
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(546..563)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1419..1448)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     RBS             1471..1493
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1508..1532
                     /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(1532..1551)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1533..1551)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1639..1658)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1864..1941
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1942..3024
                     /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(1961..1980)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3833..4024
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4018..4040)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4126..4265
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4181..4200
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4280..4297)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4451..5039)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4531..4550)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5150..5169)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5161..5976
                     /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             5161..5976
                     /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(5234..5253)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(join(6068..6128,1..395))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
       61 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
      121 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
      181 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
      241 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
      301 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
      361 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccaatcc ggatatagtt
      421 cctcctttca gcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctagt
      481 tattgctcag cggtggcagc agccaactca gcttcctttc gggctttgtt agcagccgga
      541 tctcagtggt ggtggtggtg gtgctcgagt cattacgaac cgccggtacg ttgcaagata
      601 tgaacacctg agttggtttc gatgatattg gacacttcac caacatgcag gttaaaggca
      661 gcctcttcaa atggcggctg catttgacct ttgctgaaaa aacccagatc accaccacga
      721 tcgtggctgc tgcaatcaga ttcggtattt gccagctcgc tcaatttcac ctcgccggat
      781 aagatacgtt cgaggtgctt cttcaggatc tgaatgcttt cgtcacgagt gcgagaaatg
      841 ccatccggac tcttccagct cttcggtttg cggctttggt tgtttttaat cagcaagtgc
      901 gagactctaa cctggccgtc ctcattaacc agcggcttgt aaccgttatt tttgaatttc
      961 gcgatatacg cattcagcac ctctttgtcg gtgccgtacg gcgggtccca gctggactcg
     1021 ttcgtagact ggttcaggaa atactccttg ttgtggctac ggctcacgcg aatggtccag
     1081 tttgggggta gtcctgttga agtactagcc attccagagc cgccaatctg ttcacggtgg
     1141 gcttcaatga tgtcgttgtc ttccatgtcc aggtcttccg gcgcttgatc tgcctggatg
     1201 cgaataccgt catacaggaa acgcaggcta tccatttcct tgccctgacg tttggcaaac
     1261 gcttccatca ggcgacgcag cggcgtggtc tttttgatct taaagaaaat ttcagagctg
     1321 ccgtcactaa ctttcaggtt aatatgggtt tccggtttca cttccggctt gacttccggc
     1381 tttgcttctt ggttgacttc cgaatcttgc aggctaccgt ggtgatggtg atggtgatga
     1441 tggtgatgaa tcttcgccat catatgtata tctccttctt aaagttaaac aaaattattt
     1501 ctagagggga attgttatcc gctcacaatt cccctatagt gagtcgtatt aatttcgcgg
     1561 gatcgagatc gatctcgatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca
     1621 caggtgcggt tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc
     1681 acttcgggct catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg
     1741 gactgttggg cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc
     1801 tcaacctact actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgagatc
     1861 ccggacacca tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag
     1921 agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc
     1981 ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa
     2041 acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca
     2101 caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg
     2161 cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc
     2221 gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat
     2281 cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc
     2341 attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag
     2401 acacccatca acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat
     2461 ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg
     2521 gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata
     2581 gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg
     2641 aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca
     2701 atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg acatctcggt agtgggatac
     2761 gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt
     2821 cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg
     2881 aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat
     2941 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     3001 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta
     3061 ggcaccggga tctcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg
     3121 ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt
     3181 aggacaggtg ccggcagcgc tctgggtcat tttcggcgag gaccgctttc gctggagcgc
     3241 gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt
     3301 cgtcactggt cccgccacca aacgtttcgg cgagaagcag gccattatcg ccggcatggc
     3361 ggccccacgg gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg
     3421 ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg
     3481 ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta
     3541 aagtctggaa acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg
     3601 atgctgctgg ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc
     3661 ctgagtgatt tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg
     3721 ttccagtaac cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt
     3781 catcggtatc attaccccca tgaacagaaa tcccccttac acggaggcat cagtgaccaa
     3841 acaggaaaaa accgccctta acatggcccg ctttatcaga agccagacat taacgcttct
     3901 ggagaaactc aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga
     3961 ccacgctgat gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     4021 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     4081 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     4141 gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta
     4201 ctgagagtgc accattgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     4261 tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4321 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4381 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4441 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4501 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4561 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4621 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4681 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4741 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4801 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4861 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     4921 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4981 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5041 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5101 ggattttggt catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     5161 atgagccata ttcaacggga aacgtcttgc tctaggccgc gattaaattc caacatggat
     5221 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     5281 tatcgattgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     5341 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
     5401 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     5461 atccccggga aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     5521 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     5581 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
     5641 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     5701 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     5761 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     5821 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     5881 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     5941 ttgcagtttc atttgatgct cgatgagttt ttctaagaat taattcatga gcggatacat
     6001 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6061 gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     6121 agctcatt
//
