LOCUS       sequence-207745-        6008 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of murine osteopontin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6008)
  AUTHORS   Zhang G, He B, Weber GF
  TITLE     Growth factor signaling induces metastasis genes in transformed 
            cells: molecular connection between Akt kinase and osteopontin in 
            breast cancer.
  JOURNAL   Mol Cell Biol. 2003 Sep . 23(18):6507-19.
  PUBMED    12944477
REFERENCE   2  (bases 1 to 6008)
  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: "Mol Cell 
            Biol. 2003 Sep . 23(18):6507-19."
COMMENT     Sequence Label: mOPN-PB0
FEATURES             Location/Qualifiers
     source          1..6008
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..470
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    535..734
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             935..1351
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1281..1303
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1319..1335
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             2275..2298
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2331..2351)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2336..2665
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2516..2651
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2675..3274
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2675..2694)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3171..3191
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3495..3964
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4094..4111)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4265..4853)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4345..4364)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5013..5804,5805..5873))
                     /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     5636..5655
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5874..5978)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
       61 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
      121 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
      181 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
      241 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
      301 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
      361 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
      421 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct
      481 cgtccgggat cgggagaccc ctgcccaggg accaccgacc caccaccggg aggcaagctg
      541 gccagcaact tatctgtgtc tgtccgattg tctagtgtct atgactgatt ttatgcgcct
      601 gcgtcggtac tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga
      661 gttctgaaca cccggccgca accctgggag acgtcccagg gactttgggg gccgtttttg
      721 tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat atgtggttct
      781 ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg
      841 gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt gttgtctctg
      901 tctgactgtg tttctgtatt tgtctgaaaa ttagggccag actgttacca ctcccttaag
      961 tttgacctta ggtcactgga aagatgtcga gcggatcgct cacaaccagt cggtagatgt
     1021 caagaagaga cgttgggtta ccttctgctc tgcagaatgg ccaaccttta acgtcggatg
     1081 gccgcgagac ggcaccttta accgagacct catcacccag gttaagatca aggtcttttc
     1141 acctggcccg catggacacc cagaccaggt cccctacatc gtgacctggg aagccttggc
     1201 ttttgacccc cctccctggg tcaagccctt tgtacaccct aagcctccgc ctcctcttcc
     1261 tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct
     1321 ttatccagcc ctcactcctt ctctaggcgc cggccggatc ccgaccatga gattggcagt
     1381 gatttgcttt tgcctgtttg gcattgcctc ctccctcccg gtgaaagtga ctgattctgg
     1441 cagctcagag gagaagcttt acagcctgca cccagatcct atagccacat ggctggtgcc
     1501 tgacccatct cagaagcaga atctccttgc gccacagaat gctgtgtcct ctgaagaaaa
     1561 ggatgacttt aagcaagaaa ctcttccaag caattccaat gaaagccatg accacatgga
     1621 cgacgatgat gacgatgatg atgacgatgg agaccatgca gagagcgagg attctgtgga
     1681 ctcggatgaa tctgacgaat ctcaccattc ggatgagtct gatgagaccg tcactgctag
     1741 tacacaagca gacactttca ctccaatcgt ccctacagtc gatgtcccca acggccgagg
     1801 tgatagcttg gcttatggac tgaggtcaaa gtctaggagt ttccaggttt ctgatgaaca
     1861 gtatcctgat gccacagatg aggacctcac ctctcacatg aagagcggtg agtctaagga
     1921 gtccctcgat gtcatccctg ttgcccagct tctgagcatg ccctctgatc aggacaacaa
     1981 cggaaagggc agccatgagt caagtcagct ggatgaacca agtctggaaa cacacagact
     2041 tgagcattcc aaagagagcc aggagagtgc cgatcagtcg gatgtgatcg atagtcaagc
     2101 aagttccaaa gccagcctgg aacatcagag ccacaagttt cacagccaca aggacaagct
     2161 agtcctagac cctaagagta aggaagatga taggtatctg aaattccgaa tttctcatga
     2221 attagagagt tcatcttctg aggtcaacta aagaagaggc aaaaacacag ttccgactac
     2281 aaggacgacg atgacaagtg agaattcgcc agcacagtgg tcgaccctgt ggaatgtgtg
     2341 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2401 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2461 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     2521 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     2581 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     2641 ttttggaggc ctaggctttt gcaaaaagct taccatgacc gagtacaagc ccacggtgcg
     2701 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     2761 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     2821 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     2881 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     2941 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     3001 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     3061 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     3121 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     3181 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     3241 cacctggtgc atgacccgca agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc
     3301 gaccgaaagg agcgcacgac cccatgcatc gataaaataa aagattttat ttagtctcca
     3361 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc
     3421 attttgcaag gcatggaaaa atacataact gagaatagag aagttcagat caaggtcagg
     3481 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     3541 cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt
     3601 aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc ggtccagccc
     3661 tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc
     3721 ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc
     3781 tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga
     3841 ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg
     3901 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct
     3961 ttcacatgca gcatgtatca aaattaattt ggtttttttt cttaagtatt tacattaaat
     4021 ggccatagtt gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4081 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4141 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4201 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4261 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4321 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4381 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4441 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4501 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4561 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4621 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4681 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4741 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4801 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4861 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4921 tggtcatgag attatcaaaa aggatcttca cctagatcct tttgcggccg caaatcaatc
     4981 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5041 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5101 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5161 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5221 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5281 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5341 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5401 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5461 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5521 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5581 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     5641 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5701 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5761 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5821 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5881 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5941 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6001 cctgcagc
//
