LOCUS       sequence-430368-        5991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for production of HIV p24 protein with 
            a polyhistidine tag at the N-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5991)
  TITLE     Bacterial expression plasmid for production of HIV p24 protein with 
            a polyhistidine tag at the N-terminus
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5991)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pET28a-HIV p24
FEATURES             Location/Qualifiers
     source          1..5991
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(71..90)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     281..302
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(531..1346)
                     /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(531..1346)
                     /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     1254..1273
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1338..1357
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1468..2056
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1957..1976
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2210..2227
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2242..2381
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2307..2326)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2467..2489
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2483..2674)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3483..4565)
                     /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     4527..4546
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4566..4643)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4849..4868
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4952..4971
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4952..4970
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4971..4995
                     /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             5010..5032
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5051..5068
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5078..5095
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             5807..5824
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5877..5895)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5891..5938
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      join(5975..5991,1..439)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
       61 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
      121 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
      181 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
      241 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
      301 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
      361 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
      421 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
      481 gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac
      541 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
      601 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
      661 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
      721 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
      781 gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc
      841 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
      901 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
      961 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     1021 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta
     1081 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     1141 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     1201 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
     1261 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa
     1321 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
     1381 agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     1441 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     1501 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1561 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     1621 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1681 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1741 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1801 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1861 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1921 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1981 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     2041 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     2101 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     2161 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     2221 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     2281 ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     2341 tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac
     2401 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     2461 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
     2521 agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat
     2581 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg
     2641 ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc
     2701 tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg
     2761 atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc
     2821 ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag
     2881 gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg
     2941 gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg
     3001 ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg
     3061 attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga
     3121 gcacgatcat gcgcacccgt ggggccgcca tgccggcgat aatggcctgc ttctcgccga
     3181 aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata
     3241 ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga
     3301 cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg
     3361 cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca
     3421 agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc
     3481 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     3541 aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt
     3601 gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg
     3661 tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata
     3721 taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc
     3781 agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc
     3841 atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg
     3901 gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta
     3961 tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg
     4021 atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg
     4081 gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca
     4141 ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc
     4201 agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg
     4261 cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc
     4321 gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc
     4381 aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc
     4441 atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg
     4501 cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt
     4561 ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag
     4621 gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat
     4681 taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc
     4741 atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
     4801 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
     4861 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
     4921 gtagaggatc gagatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga
     4981 gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga gatataccat
     5041 gggcagcagc catcatcatc atcatcacag cagcggcctg gtgccgcgcg gcagccatat
     5101 gcctattgtg caaaaccttc aagggcaaat ggtccaccag gcaatttccc caagaacatt
     5161 aaatgcgtgg gtcaaggttg ttgaagagaa ggcattctcc ccggaagtga tacccatgtt
     5221 ctcggcttta tccgagggag ctactccaca ggaccttaac acgatgctta atacggtagg
     5281 gggccatcag gctgcgatgc agatgttgaa ggaaactata aacgaagagg ccgcagagtg
     5341 ggatcggctt caccccgtcc acgcgggacc aatagcaccc ggtcagatgc gggaaccgag
     5401 aggatccgac atagcaggta ctacgagtac cttacaggaa cagatagggt ggatgaccca
     5461 caatcctcca attcccgtag gggagatata caagagatgg atcattttag gactgaataa
     5521 gatagtccgt atgtattccc ccacgtctat tttggatatc cgtcaaggtc cgaaagagcc
     5581 ttttcgtgat tacgttgacc gtttctacaa aacccttcgt gcagaacaag ccagccagga
     5641 agtaaagaat tggatgactg aaacgttgtt ggttcagaat gcaaaccccg attgtaaaac
     5701 tatcctgaag gctctgggtc ccggtgcaac cttggaagaa atgatgacag cgtgccaagg
     5761 cgtcggcgga cccggccaca aagcacgggt tctgtaataa ctcgagcacc accaccacca
     5821 ccactgagat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     5881 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
     5941 gaaaggagga actatatccg gattggcgaa tgggacgcgc cctgtagcgg c
//
