LOCUS       sequence-207738-        6076 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 6076)
  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 6076)
  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 M5
FEATURES             Location/Qualifiers
     source          1..6076
                     /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(1814..1831)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1820..2044
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2176..2764)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2256..2275)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3093..3140
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3165..3184
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3372..4166)
                     /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(3372..4166)
                     /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(3483..3502)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4093..4112
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4201..4558)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4215..4350
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4367..4558
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4515..4535
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(4618..5409,5410..5478))
                     /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     5241..5260
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5479..5583)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5610..6065
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5697..5716)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5907..5928
                     /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 aaaagcaggc taatgctgtg tcctctgaag aaaccaatga ctttaaacaa
      901 gagacccttc caagtaagtc caacgaaagc catgaccaca tggatgatat ggatgatgaa
      961 gatgatgacg accatgtgga cagccaggac tccattgact cgaacgactc tgatgatgta
     1021 gatgacactg atgattctca ccagtctgat gagtctcacc attctgatga atctgatgaa
     1081 ctggtcactg attttcccac ggacctgcca gcaaccgaag ttttcactcc agttgtcccc
     1141 acagtagaca catatgatgg ccgaggtgat agtgtggttt atggactgag gtcaaaatct
     1201 aagaagtttc gcagacctga catccagtac cctgatgcta cagacgagga catcacctca
     1261 cacatggaaa gcgaggagtt gaatggtgca tacaaggcca tccccgttgc ccaggacctg
     1321 aacgcgcctt ctgattggga cagccgtggg aaggacagtt atgaaacgag tcagctggat
     1381 gaccagagtg ctgaaaccca cagccacaag cagtccagat tatataagcg gaaagctaat
     1441 gatgagagca atgagcattc cgatgtgatt gatagtcagg aactttccaa agtcagccgt
     1501 gaattccaca gccatgaatt tcacagccat gaagatatgc tggttgtaga ccccaaaagt
     1561 aaggaagaag ataaacacct gaaatttcgt atttctcatg aattagatag tgcatcttct
     1621 gaggtcaatt aaaaggagaa aaaatacaat ttctcacttt gcatttagtc aaaagaaaaa
     1681 atgctttata gcaaaatgaa agagaacatg aaatgcttct ttctcagttt attggttgaa
     1741 gccgaattct gcagatatcc atcacactgg cggccgctcg agtctagagg gcccgtttaa
     1801 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     1861 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     1921 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     1981 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     2041 atggcttctg aggcggaaag aaccagtggc ggtaatacgg ttatccacag aatcagggga
     2101 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2161 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2221 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2281 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2341 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2401 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2461 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2521 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2581 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2641 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2701 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2761 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2821 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2881 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa cctgaggcta tggcagggcc
     2941 tgccgccccg acgttggctg cgagccctgg gccttcaccc gaacttgggg ggtggggtgg
     3001 ggaaaaggaa gaaacgcggg cgtattggcc ccaatggggt ctcggtgggg tatcgacaga
     3061 gtgccagccc tgggaccgaa ccccgcgttt atgaacaaac gacccaacac cgtgcgtttt
     3121 attctgtctt tttattgccg tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt
     3181 ttcagttagc ctccccctag ggtgggcgaa gaactccagc atgagatccc cgcgctggag
     3241 gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc
     3301 ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc
     3361 cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg
     3421 ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca
     3481 gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca
     3541 cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg
     3601 ccatgggtca cgacgagatc ctcgccgtcg ggcatgctcg ccttgagcct ggcgaacagt
     3661 tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct
     3721 tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta
     3781 gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca
     3841 ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc
     3901 cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc
     3961 cacgatagcc gcgctgcctc gtcttgcagt tcattcaggg caccggacag gtcggtcttg
     4021 acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg
     4081 attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct
     4141 gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcgatc
     4201 tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc
     4261 cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg
     4321 ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc
     4381 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     4441 acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     4501 tggggacttt ccacacccta actgacacac attccacagc tggttctttc cgcctcagga
     4561 ctcttccttt ttcaataaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4621 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4681 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4741 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4801 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4861 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4921 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4981 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5041 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5101 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5161 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5221 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5281 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5341 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5401 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5461 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5521 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5581 gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc
     5641 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     5701 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     5761 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     5821 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     5881 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     5941 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     6001 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     6061 aatttacgcg cgcgtt
//
