LOCUS       sequence-464156-        6041 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Heterologous protein expression of YqeY in Escherichia coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6041)
  TITLE     AI Proteins, Inc. plasmids
REFERENCE   2  (bases 1 to 6041)
  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_1NG6
FEATURES             Location/Qualifiers
     source          1..6041
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             374..565
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(559..581)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    667..806
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     722..741
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(821..838)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(992..1580)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1072..1091)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1691..1710)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1702..2517
                     /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             1702..2517
                     /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(1775..1794)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(2609..3064)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2746..2767)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2958..2977
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(3101..3148)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3144..3162
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(3215..3232)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4001..4030)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     RBS             4053..4075
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4090..4114
                     /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(4114..4133)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4115..4133)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4221..4240)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        4446..4523
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4524..5606
                     /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(4543..4562)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt
       61 cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct gcaccattat
      121 gttccggatc tgcatcgcag gatgctgctg gctaccctgt ggaacaccta catctgtatt
      181 aacgaagcgc tggcattgac cctgagtgat ttttctctgg tcccgccgca tccataccgc
      241 cagttgttta ccctcacaac gttccagtaa ccgggcatgt tcatcatcag taacccgtat
      301 cgtgagcatc ctctctcgtt tcatcggtat cattaccccc atgaacagaa atccccctta
      361 cacggaggca tcagtgacca aacaggaaaa aaccgccctt aacatggccc gctttatcag
      421 aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg aacaggcaga
      481 catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc tcgcgcgttt
      541 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
      601 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
      661 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat
      721 gcggcatcag agcagattgt actgagagtg caccattgcg gtgtgaaata ccgcacagat
      781 gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc
      841 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
      901 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
      961 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     1021 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     1081 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     1141 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     1201 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     1261 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     1321 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     1381 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     1441 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     1501 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     1561 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     1621 ggaacgaaaa ctcacgttaa gggattttgg tcatgaacaa taaaactgtc tgcttacata
     1681 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcttg ctctaggccg
     1741 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     1801 gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt
     1861 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
     1921 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     1981 gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt agaagaatat
     2041 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     2101 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
     2161 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
     2221 cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc
     2281 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     2341 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     2401 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     2461 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaagaa
     2521 ttaattcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     2581 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
     2641 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     2701 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     2761 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     2821 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     2881 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     2941 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     3001 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     3061 gcgtcccatt cgccaatccg gatatagttc ctcctttcag caaaaaaccc ctcaagaccc
     3121 gtttagaggc cccaaggggt tatgctagtt attgctcagc ggtggcagca gccaactcag
     3181 cttcctttcg ggctttgtta gcagccggat ctcagtggtg gtggtggtgg tgctcgagtc
     3241 attacgaacc ggacagctga gagctgacca atttgttaat caggctacca tcagccttac
     3301 ccttgacttt cggcataatt gcgcccatca ccttgcccat gtcagctttg gacgacgcac
     3361 caacctccgc aatggtttca ttaacaatgg tacgcagctc ctcttcagac agctgttccg
     3421 gcaggtacac ttccaggatg tcgagttctt tctgcacctt atcaaccagg tccaggcgat
     3481 tcgcgttgct gaactcttgc aggctgtctt tacgttgttt cagctcacgg ctcagaacgg
     3541 tcaactcttc atcttcagtc aagctatctt tcttaagttt gatcgcctcg ttttgcaagg
     3601 acgccttaac catacgcacc accgttaatt tgtccttctc gcggttcttc atatacagct
     3661 tcatatcttg attcaacctt tctaatagtg acattccaga gccgccaatc tgttcacggt
     3721 gggcttcaat gatgtcgttg tcttccatgt ccaggtcttc cggcgcttga tctgcctgga
     3781 tgcgaatacc gtcatacagg aaacgcaggc tatccatttc cttgccctga cgtttggcaa
     3841 acgcttccat caggcgacgc agcggcgtgg tctttttgat cttaaagaaa atttcagagc
     3901 tgccgtcact aactttcagg ttaatatggg tttccggttt cacttccggc ttgacttccg
     3961 gctttgcttc ttggttgact tccgaatctt gcaggctacc gtggtgatgg tgatggtgat
     4021 gatggtgatg aatcttcgcc atcatatgta tatctccttc ttaaagttaa acaaaattat
     4081 ttctagaggg gaattgttat ccgctcacaa ttcccctata gtgagtcgta ttaatttcgc
     4141 gggatcgaga tcgatctcga tcctctacgc cggacgcatc gtggccggca tcaccggcgc
     4201 cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg
     4261 ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg
     4321 gggactgttg ggcgccatct ccttgcatgc accattcctt gcggcggcgg tgctcaacgg
     4381 cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag agcgtcgaga
     4441 tcccggacac catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag
     4501 agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg
     4561 ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga
     4621 aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg
     4681 cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc
     4741 tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca
     4801 gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca
     4861 atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg
     4921 ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc
     4981 agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc
     5041 atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct
     5101 cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga
     5161 tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc
     5221 tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg
     5281 caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggacatctcg gtagtgggat
     5341 acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt
     5401 ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg
     5461 tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca
     5521 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     5581 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat
     5641 taggcaccgg gatctcgacc gatgcccttg agagccttca acccagtcag ctccttccgg
     5701 tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc
     5761 gtaggacagg tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc
     5821 gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc
     5881 ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg
     5941 gcggccccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg
     6001 ggttgcctta ctggttagca gaatgaatca ccgatacgcg a
//
