LOCUS       sequence-464151-        6047 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Heterologous protein expression of rpb7-binding protein seb1 in 
            Escherichia coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6047)
  TITLE     AI Proteins, Inc. plasmids
REFERENCE   2  (bases 1 to 6047)
  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_5MDT
FEATURES             Location/Qualifiers
     source          1..6047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     702..721
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     786..805
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      916..1504
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1405..1424
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1658..1675
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1690..1829
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1755..1774)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1915..1937
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1931..2122)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2931..4013)
                     /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     3975..3994
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4014..4091)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4297..4316
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4404..4423
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4404..4422
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4423..4447
                     /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             4462..4484
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4507..4536
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             5311..5328
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5381..5399)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5395..5442
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      5479..5934
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5566..5585)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5776..5797
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(6026..6047,1..794))
                     /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(6026..6047,1..794))
                     /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 aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt
       61 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
      121 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
      181 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
      241 agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
      301 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg
      361 cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact
      421 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct
      481 gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
      541 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
      601 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
      661 ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
      721 ccatataaat cagcatccat gttggaattt aatcgcggcc tagagcaaga cgtttcccgt
      781 tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt
      841 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
      901 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
      961 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1021 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     1081 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     1141 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     1201 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     1261 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     1321 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     1381 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     1441 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     1501 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     1561 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     1621 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     1681 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa
     1741 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     1801 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     1861 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     1921 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     1981 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     2041 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     2101 cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg
     2161 taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg
     2221 cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga
     2281 gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg
     2341 gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc
     2401 gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg
     2461 cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct
     2521 aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc
     2581 gcacccgtgg ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg
     2641 cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca
     2701 ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg
     2761 ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag
     2821 tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc
     2881 gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg
     2941 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3001 gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac
     3061 agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt
     3121 tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg
     3181 tcttcggtat cgtcgtatcc cactaccgag atgtccgcac caacgcgcag cccggactcg
     3241 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
     3301 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
     3361 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
     3421 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
     3481 cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata
     3541 ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca
     3601 gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg
     3661 cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc
     3721 atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt
     3781 tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg
     3841 cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc
     3901 actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc
     3961 tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc
     4021 accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat
     4081 tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc
     4141 cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat
     4201 ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct
     4261 catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc
     4321 agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga
     4381 gatcgatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
     4441 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatataca tatgatggcg
     4501 aagattcatc accatcatca ccatcaccat caccacggta gcctgcaaga ttcggaagtc
     4561 aaccaagaag caaagccgga agtcaagccg gaagtgaaac cggaaaccca tattaacctg
     4621 aaagttagtg acggcagctc tgaaattttc tttaagatca aaaagaccac gccgctgcgt
     4681 cgcctgatgg aagcgtttgc caaacgtcag ggcaaggaaa tggatagcct gcgtttcctg
     4741 tatgacggta ttcgcatcca ggcagatcaa gcgccggaag acctggacat ggaagacaac
     4801 gacatcattg aagcccaccg tgaacagatt ggcggctctg gatcagggat tgctgaattt
     4861 gatggaatac tagactcctt agagcacagc aaaactggta ttagcggcag caagatcctc
     4921 aagttgacga acctgagcat ggaaaacgtg agcgagaacg cacagtttgt ggcttctgtt
     4981 tataaatacg ccaagcgcgc accggtcacc cataaactgg gcgcgctgta catcctggat
     5041 agcattgtga gatcattcca agatggtgct aaaaagaata atgaatcgtt cgagaacccg
     5101 gttgatgcgt cgttcagcgg cggttggtgt aaagcggcag aaattaccga ctctctggtt
     5161 gctgatgcga ttcagcatgc gccgagcgcg cacctgccga aaatcttgaa gctttgcgac
     5221 atctgggaaa aagcgtccac ctttccaccg gagaagttgg agagcctgcg ttccaagctg
     5281 aaggacgcca tgggttcgta atgactcgag caccaccacc accaccactg agatccggct
     5341 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
     5401 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
     5461 tccggattgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     5521 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     5581 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     5641 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     5701 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     5761 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     5821 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     5881 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     5941 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     6001 tatgtatccg ctcatgaatt aattcttaga aaaactcatc gagcatc
//
