LOCUS       sequence-444263-        5771 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of recombinant BMPS WT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5771)
  TITLE     Abu-Remaileh Lab Plasmids 2023
REFERENCE   2  (bases 1 to 5771)
  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: pRK5 BMPS WT-6xHis
FEATURES             Location/Qualifiers
     source          1..5771
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     828..845
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     859..878
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             2017..2034
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      2071..2080
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2084..2218
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2121..2140)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2175..2194
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2287..2644
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2287..2478
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2310..2330)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2495..2630
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2664..2681)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2664..2680)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2673..2695)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2893..3348
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2980..2999)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3190..3211
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3365..3384)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3484..3506
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3544..3562)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3630..3734
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3735..3803,3804..4595)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3953..3972)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4766..5354
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5255..5274
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5508..5525
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5642..5663
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5678..5683,5684..5701,5702..5708)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5716..5732
                     /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     5721..5743
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5740..5756
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5740..5756
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc tatcgattga attcaccatg gcgcaggagg tagacacggc acagggcgcc
      961 gagatgcggc ggggcgcggg cgcggctcgg ggacgcgctt cctggtgctg ggccctggcg
     1021 ctgctttggc tcgcggtggt tccgggctgg tcccgggtct cgggcatccc ctcccggcgc
     1081 cactggccgg tgccctacaa gcgctttgac ttccgtccaa aacctgatcc ttattgtcaa
     1141 gctaagtata ctttctgtcc aactggctca cctatcccag ttatggaggg tgatgatgac
     1201 attgaagttt ttcgattaca agccccagta tgggaattta aatatggaga cctcctggga
     1261 cacttgaaaa ttatgcatga tgccattgga ttcagaagta cattaactgg caagaactac
     1321 acaatggaat ggtatgaact tttccaactt ggcaactgta catttcccca tctccgacct
     1381 gaaatggatg cccctttctg gtgtaatcaa ggcgctgcct gcttttttga gggaattgat
     1441 gatgttcact ggaaggaaaa tgggacatta gttcaagtag caactatatc aggaaacatg
     1501 ttcaaccaaa tggcaaagtg ggtgaaacag gacaatgaaa caggaattta ttatgagaca
     1561 tggaatgtaa aagccagccc agaaaagggg gcagagacat ggtttgattc ctacgactgt
     1621 tccaaatttg tgttaaggac ctttaacaag ttggctgaat ttggagcaga gttcaagaac
     1681 atagaaacca actatacaag aatatttctt tacagtggag aacctactta tctgggaaat
     1741 gaaacatctg tttttgggcc aacaggaaac aagactcttg gtttagccat aaaaagattt
     1801 tattacccct tcaaaccaca tttgccaact aaagaatttc tgttgagtct cttgcaaatt
     1861 tttgatgcag tgattgtgca caaacagttc tatttgtttt ataattttga atattggttt
     1921 ttacctatga aattcccttt tattaaaata acatatgaag aaatcccttt acctatcaga
     1981 aacaaaacac tctctggttt aggaggtgga ggtagtcatc atcaccatca ccactaggcg
     2041 gccgctaagt aagtaaggat ccccagcttg gccgccatgg cccaacttgt ttattgcagc
     2101 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     2161 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcgg
     2221 gaattaattc ggcgcagcac catggcctga aataacctct gaaagaggaa cttggttagg
     2281 taccttctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     2341 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     2401 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2461 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     2521 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     2581 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     2641 caaaaagctg ttaacagctt ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     2701 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     2761 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     2821 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     2881 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2941 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     3001 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3061 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     3121 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3181 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcgggctatt
     3241 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     3301 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     3361 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     3421 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     3481 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     3541 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     3601 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3661 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3721 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3781 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3841 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3901 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3961 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4021 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     4081 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4141 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4201 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4261 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     4321 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4381 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4441 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4501 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4561 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4621 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4681 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4741 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4801 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4861 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4921 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4981 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5041 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5101 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5161 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5221 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5281 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5341 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5401 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     5461 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     5521 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     5581 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     5641 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     5701 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgacatga
     5761 ttacgaatta a
//
