LOCUS       sequence-207737-        6070 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mutant osteopontin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6070)
  AUTHORS   Shen H, Weber GF
  TITLE     The osteopontin-c splice junction is important for 
            anchorage-independent growth.
  JOURNAL   Mol Carcinog. 2014 Jun;53(6):480-7. doi: 10.1002/mc.21997. Epub 2013
            Jan 28.
  PUBMED    23359517
REFERENCE   2  (bases 1 to 6070)
  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"; doi: "10.1002/mc.21997"; 
            journalName: "Mol Carcinog"; date: "2014-06"; volume: "53"; issue: 
            "6"; pages: "480-7"
COMMENT     Sequence Label: pCR3.1-OPNc M4
FEATURES             Location/Qualifiers
     source          1..6070
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1808..1825)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1814..2038
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2170..2758)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2250..2269)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3087..3134
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3159..3178
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3366..4160)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(3366..4160)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3477..3496)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4087..4106
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4195..4552)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4209..4344
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4361..4552
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4509..4529
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(4612..5403,5404..5472))
                     /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     5235..5254
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5473..5577)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5604..6059
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5691..5710)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5901..5922
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc actagtaacg gccgccagtg
      721 tgctggaatt cggcttcaaa cgccgaccaa ggaaaactca ctaccatgag aattgcagtg
      781 atttgctttt gcctcctagg catcacctgt gccataccag ttaaacaggc tgattctgga
      841 agttctgagg aaaagaatgc tgtgtcctct gaagaaacca atgactttaa acaagagacc
      901 cttccaagta agtccaacga aagccatgac cacatggatg atatggatga tgaagatgat
      961 gacgaccatg tggacagcca ggactccatt gactcgaacg actctgatga tgtagatgac
     1021 actgatgatt ctcaccagtc tgatgagtct caccattctg atgaatctga tgaactggtc
     1081 actgattttc ccacggacct gccagcaacc gaagttttca ctccagttgt ccccacagta
     1141 gacacatatg atggccgagg tgatagtgtg gtttatggac tgaggtcaaa atctaagaag
     1201 tttcgcagac ctgacatcca gtaccctgat gctacagacg aggacatcac ctcacacatg
     1261 gaaagcgagg agttgaatgg tgcatacaag gccatccccg ttgcccagga cctgaacgcg
     1321 ccttctgatt gggacagccg tgggaaggac agttatgaaa cgagtcagct ggatgaccag
     1381 agtgctgaaa cccacagcca caagcagtcc agattatata agcggaaagc taatgatgag
     1441 agcaatgagc attccgatgt gattgatagt caggaacttt ccaaagtcag ccgtgaattc
     1501 cacagccatg aatttcacag ccatgaagat atgctggttg tagaccccaa aagtaaggaa
     1561 gaagataaac acctgaaatt tcgtatttct catgaattag atagtgcatc ttctgaggtc
     1621 aattaaaagg agaaaaaata caatttctca ctttgcattt agtcaaaaga aaaaatgctt
     1681 tatagcaaaa tgaaagagaa catgaaatgc ttctttctca gtttattggt tgaagccgaa
     1741 ttctgcagat atccatcaca ctggcggccg ctcgagtcta gagggcccgt ttaaacccgc
     1801 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     1861 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     1921 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     1981 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     2041 tctgaggcgg aaagaaccag tggcggtaat acggttatcc acagaatcag gggataacgc
     2101 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2161 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2221 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2281 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2341 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2401 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2461 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2521 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2581 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2641 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2701 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2761 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     2821 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     2881 aagttttaaa tcaatctaaa gtatatatga gtaacctgag gctatggcag ggcctgccgc
     2941 cccgacgttg gctgcgagcc ctgggccttc acccgaactt ggggggtggg gtggggaaaa
     3001 ggaagaaacg cgggcgtatt ggccccaatg gggtctcggt ggggtatcga cagagtgcca
     3061 gccctgggac cgaaccccgc gtttatgaac aaacgaccca acaccgtgcg ttttattctg
     3121 tctttttatt gccgtcatag cgcgggttcc ttccggtatt gtctccttcc gtgtttcagt
     3181 tagcctcccc ctagggtggg cgaagaactc cagcatgaga tccccgcgct ggaggatcat
     3241 ccagccggcg tcccggaaaa cgattccgaa gcccaacctt tcatagaagg cggcggtgga
     3301 atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga accccagagt
     3361 cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg
     3421 gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata
     3481 tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg
     3541 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg
     3601 gtcacgacga gatcctcgcc gtcgggcatg ctcgccttga gcctggcgaa cagttcggct
     3661 ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc
     3721 cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga
     3781 tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca
     3841 aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc
     3901 gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat
     3961 agccgcgctg cctcgtcttg cagttcattc agggcaccgg acaggtcggt cttgacaaaa
     4021 agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc
     4081 tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc
     4141 aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc gatctttgca
     4201 aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc
     4261 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
     4321 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
     4381 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct
     4441 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     4501 ctttccacac cctaactgac acacattcca cagctggttc tttccgcctc aggactcttc
     4561 ctttttcaat aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4621 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4681 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4741 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4801 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4861 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4921 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4981 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5041 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5101 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5161 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5221 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5281 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5341 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5401 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5461 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5521 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5581 atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     5641 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     5701 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     5761 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     5821 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     5881 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     5941 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     6001 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     6061 cgcgcgcgtt
//
