LOCUS       sequence-464124-        5918 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Heterologous protein expression of YfiB in Escherichia coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5918)
  TITLE     AI Proteins, Inc. plasmids
REFERENCE   2  (bases 1 to 5918)
  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_4ZHW
FEATURES             Location/Qualifiers
     source          1..5918
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     368..387
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     452..471
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      582..1170
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1071..1090
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1324..1341
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1356..1495
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1421..1440)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1581..1603
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1597..1788)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2597..3679)
                     /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     3641..3660
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3680..3757)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3963..3982
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4070..4089
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4070..4088
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4089..4113
                     /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)."
     RBS             4128..4150
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4173..4202
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             4848..4865
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4918..4936)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4932..4979
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      5016..5471
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5103..5122)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5313..5334
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(5563..5918,1..460))
                     /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(join(5563..5918,1..460))
                     /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"
ORIGIN
        1 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
       61 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
      121 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
      181 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
      241 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
      301 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
      361 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
      421 gcggcctaga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
      481 ttatgtaagc agacagtttt attgttcatg accaaaatcc cttaacgtga gttttcgttc
      541 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
      601 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
      661 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
      721 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
      781 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
      841 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
      901 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
      961 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     1021 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     1081 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     1141 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     1201 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     1261 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     1321 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     1381 catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg
     1441 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
     1501 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     1561 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     1621 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     1681 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     1741 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt
     1801 gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac
     1861 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
     1921 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt
     1981 taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa
     2041 cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa
     2101 gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg
     2161 ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt
     2221 cctcaacgac aggagcacga tcatgcgcac ccgtggggcc gccatgccgg cgataatggc
     2281 ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg
     2341 caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc
     2401 ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa
     2461 gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg
     2521 gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac
     2581 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     2641 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt
     2701 ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga
     2761 gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg
     2821 ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatgt
     2881 ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat
     2941 cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt
     3001 gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc
     3061 gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc
     3121 ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg
     3181 taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa
     3241 ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg
     3301 gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac
     3361 aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg
     3421 cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg
     3481 caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt
     3541 aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg
     3601 cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt
     3661 ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg
     3721 ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta
     3781 tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc
     3841 gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
     3901 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
     3961 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
     4021 acgatgcgtc cggcgtagag gatcgagatc gatctcgatc ccgcgaaatt aatacgactc
     4081 actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt
     4141 aagaaggaga tatacatatg atggcgaaga ttcatcacca tcatcaccat caccatcacc
     4201 acggtagcct gcaagattcg gaagtcaacc aagaagcaaa gccggaagtc aagccggaag
     4261 tgaaaccgga aacccatatt aacctgaaag ttagtgacgg cagctctgaa attttcttta
     4321 agatcaaaaa gaccacgccg ctgcgtcgcc tgatggaagc gtttgccaaa cgtcagggca
     4381 aggaaatgga tagcctgcgt ttcctgtatg acggtattcg catccaggca gatcaagcgc
     4441 cggaagacct ggacatggaa gacaacgaca tcattgaagc ccaccgtgaa cagattggcg
     4501 gctctggaat gagttcaaaa gtactatttg gaaacaattt agaccgtttg aatccggata
     4561 gccgtaatac cctgaccaag atcgcccgtg cgctgctggc ggtggacatc gacaaagtac
     4621 gtttggaggg ccacacggac aactatggtg atgaaggcta caaccagaaa ctgtctgagc
     4681 gcagagcgga aagcgttgcg gctgtgttcc gcgaggcagg catgccggca gcgaatattg
     4741 aagttcgcgg tctgggtatg agcaagccgg ttgccgataa caaaacccgt gctggtcgtt
     4801 ccgagaaccg tcgcgtggcg attatcgtcg gttcgtaatg actcgagcac caccaccacc
     4861 accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     4921 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     4981 tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag
     5041 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     5101 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     5161 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     5221 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     5281 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     5341 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     5401 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     5461 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5521 ttctaaatac attcaaatat gtatccgctc atgaattaat tcttagaaaa actcatcgag
     5581 catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag
     5641 ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg
     5701 gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc
     5761 aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg
     5821 caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc
     5881 aaaatcactc gcatcaacca aaccgttatt cattcgtg
//
