LOCUS       sequence-464163-        5924 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Heterologous protein expression of bromodomain-containing protein 2 
            in Escherichia coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5924)
  TITLE     AI Proteins, Inc. plasmids
REFERENCE   2  (bases 1 to 5924)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pAIP4_5HFQ
FEATURES             Location/Qualifiers
     source          1..5924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      14..61
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      98..553
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(185..204)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     395..416
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(645..1460)
                     /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(645..1460)
                     /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     1368..1387
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1452..1471
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1582..2170
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2071..2090
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2324..2341
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2356..2495
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2421..2440)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2581..2603
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2597..2788)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3597..4679)
                     /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     4641..4660
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4680..4757)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4963..4982
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5070..5089
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5070..5088
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5089..5113
                     /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             5128..5150
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5173..5202
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             5854..5871
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(join(5924,1..18))
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
       61 gctgaaagga ggaactatat ccggattggc gaatgggacg cgccctgtag cggcgcatta
      121 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
      181 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
      241 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
      301 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
      361 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
      421 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
      481 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
      541 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
      601 ttttctaaat acattcaaat atgtatccgc tcatgaatta attcttagaa aaactcatcg
      661 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa
      721 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc
      781 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg
      841 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
      901 ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
      961 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga
     1021 aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg
     1081 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg
     1141 aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata
     1201 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca
     1261 tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg
     1321 ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat
     1381 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctaga gcaagacgtt
     1441 tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt
     1501 attgttcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     1561 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     1621 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     1681 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     1741 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     1801 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     1861 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     1921 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     1981 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     2041 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     2101 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     2161 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     2221 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     2281 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     2341 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     2401 ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
     2461 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
     2521 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     2581 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
     2641 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
     2701 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
     2761 taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca
     2821 tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg
     2881 aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg
     2941 accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc
     3001 cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg
     3061 acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc
     3121 aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat
     3181 tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga
     3241 tcatgcgcac ccgtggggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt
     3301 tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa
     3361 gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga
     3421 gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga
     3481 cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca
     3541 tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac
     3601 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     3661 cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg
     3721 ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg
     3781 ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat
     3841 gagctgtctt cggtatcgtc gtatcccact accgagatgt ccgcaccaac gcgcagcccg
     3901 gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca
     3961 gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc
     4021 cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag
     4081 ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc
     4141 tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa
     4201 ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg
     4261 caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca
     4321 ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt
     4381 tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg
     4441 acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac
     4501 tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc
     4561 gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa
     4621 acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca
     4681 ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg
     4741 cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa
     4801 gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa
     4861 ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca
     4921 agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata
     4981 ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag
     5041 gatcgagatc gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc
     5101 ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga tatacatatg
     5161 atggcgaaga ttcatcacca tcatcaccat caccatcacc acggtagcct gcaagattcg
     5221 gaagtcaacc aagaagcaaa gccggaagtc aagccggaag tgaaaccgga aacccatatt
     5281 aacctgaaag ttagtgacgg cagctctgaa attttcttta agatcaaaaa gaccacgccg
     5341 ctgcgtcgcc tgatggaagc gtttgccaaa cgtcagggca aggaaatgga tagcctgcgt
     5401 ttcctgtatg acggtattcg catccaggca gatcaagcgc cggaagacct ggacatggaa
     5461 gacaacgaca tcattgaagc ccaccgtgaa cagattggcg gctctggact atcagaacaa
     5521 ttaaaacact gtaatggaat cctgaaagag ttgctgagca aaaagcacgc ggcgtatgct
     5581 tggccgtttt ataaaccggt tgatgcgtct gcactgggtc tgcacgacta ccatgacatc
     5641 attaaacacc cgatggattt gtccaccgtt aagcgcaaga tggaaaaccg tgattatcgt
     5701 gacgcgcagg agttcgccgc ggatgtgcgt ctgatgttca gcaactgcta caagtacaat
     5761 ccgccagatc atgacgtcgt ggccatggca cgtaagcttc acgacgtttt tgaattccgc
     5821 tacgctaaaa tgccgggttc gtaatgactc gagcaccacc accaccacca ctgagatccg
     5881 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccac
//
