LOCUS       sequence-210255-        6154 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of full-length OPN.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6154)
  AUTHORS   He B, Mirza M, Weber GF
  TITLE     An osteopontin splice variant induces anchorage independence in 
            human breast cancer cells.
  JOURNAL   Oncogene. 2006 Apr 6;25(15):2192-202. doi: 10.1038/sj.onc.1209248.
  PUBMED    16288209
REFERENCE   2  (bases 1 to 6154)
  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: "10.1038/sj.onc"; 
            journalName: "Oncogene"; date: "2006-04-6- 6"; volume: "25"; issue: 
            "15"; pages: "2192-202"
COMMENT     Sequence Label: pCR3.1-OPNa
FEATURES             Location/Qualifiers
     source          1..6154
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        10..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     544..564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     638..657
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        638..656
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1901..1918)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1907..2131
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2263..2851)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2343..2362)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3180..3227
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3252..3271
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3459..4253)
                     /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(3459..4253)
                     /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(3570..3589)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4180..4199
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4288..4645)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4302..4437
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4454..4645
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4602..4622
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(4705..5496,5497..5565))
                     /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     5328..5347
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5566..5670)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5697..6152
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5784..5803)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5994..6015
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gcgcgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
       61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      121 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      181 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
      601 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
      661 cccaagctgg ctagcgttta aacttaagct tggtaccgag ctcggatcca ctagtaacgg
      721 ccgccagtgt gctggaattc ggcttcaaac gccgaccaag gaaaactcac taccatgaga
      781 attgcagtga tttgcttttg cctcctaggc atcacctgtg ccataccagt taaacaggct
      841 gattctggaa gttctgagga aaagcagctt tacaacaaat acccagatgc tgtggccaca
      901 tggctaaacc ctgacccatc tcagaagcag aatctcctag ccccacagaa tgctgtgtcc
      961 tctgaagaaa ccaatgactt taaacaagag acccttccaa gtaagtccaa cgaaagccat
     1021 gaccacatgg atgatatgga tgatgaagat gatgacgacc atgtggacag ccaggactcc
     1081 attgactcga acgactctga tgatgtagat gacactgatg attctcacca gtctgatgag
     1141 tctcaccatt ctgatgaatc tgatgaactg gtcactgatt ttcccacgga cctgccagca
     1201 accgaagttt tcactccagt tgtccccaca gtagacacat atgatggccg aggtgatagt
     1261 gtggtttatg gactgaggtc aaaatctaag aagtttcgca gacctgacat ccagtaccct
     1321 gatgctacag acgaggacat cacctcacac atggaaagcg aggagttgaa tggtgcatac
     1381 aaggccatcc ccgttgccca ggacctgaac gcgccttctg attgggacag ccgtgggaaa
     1441 gacagctatg aaacgagtca gctggatgac cagagtgctg aaacccacag ccacaagcag
     1501 tccagattat ataagcggaa agctaatgat gagagcaatg agcattccga tgtgattgat
     1561 agtcaggaac tttccaaagt cagccgtgaa ttccacagcc atgaatttca cagccatgaa
     1621 gatatgctgg ttgtagaccc caaaagtaag gaagaggata aacacctgaa atttcgtatt
     1681 tctcatgaat tagatagtgc atcttctgag gtcaattaaa aggagaaaaa atacaatttc
     1741 tcactttgca tttagtcaaa agaaaaaatg ctttatagca aaatgaaaga gaacatgaaa
     1801 tgcttctttc tcagtttatt ggttgaagcc gaattctgca gatatccatc acactggcgg
     1861 ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     1921 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     1981 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     2041 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     2101 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagtggcggt
     2161 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2221 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2281 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2341 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2401 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2461 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2521 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2581 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2641 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2701 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2761 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2821 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2881 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2941 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3001 tgagtaacct gaggctatgg cagggcctgc cgccccgacg ttggctgcga gccctgggcc
     3061 ttcacccgaa cttggggggt ggggtgggga aaaggaagaa acgcgggcgt attggcccca
     3121 atggggtctc ggtggggtat cgacagagtg ccagccctgg gaccgaaccc cgcgtttatg
     3181 aacaaacgac ccaacaccgt gcgttttatt ctgtcttttt attgccgtca tagcgcgggt
     3241 tccttccggt attgtctcct tccgtgtttc agttagcctc cccctagggt gggcgaagaa
     3301 ctccagcatg agatccccgc gctggaggat catccagccg gcgtcccgga aaacgattcc
     3361 gaagcccaac ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg
     3421 cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc agaagaactc gtcaagaagg
     3481 cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg
     3541 tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga
     3601 tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc
     3661 accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc
     3721 atgctcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc
     3781 agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt
     3841 ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca
     3901 tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc
     3961 ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct
     4021 gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttgcagttca
     4081 ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc
     4141 cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc
     4201 ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac
     4261 gatcctcatc ctgtctcttg atcgatcttt gcaaaagcct aggcctccaa aaaagcctcc
     4321 tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa
     4381 aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg
     4441 cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct
     4501 gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct
     4561 ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt
     4621 ccacagctgg ttctttccgc ctcaggactc ttcctttttc aataaatcaa tctaaagtat
     4681 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4741 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4801 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4861 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4921 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4981 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5041 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5101 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5161 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5221 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5281 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5341 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5401 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5461 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5521 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5581 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     5641 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc
     5701 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     5761 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     5821 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     5881 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     5941 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     6001 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     6061 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     6121 gaattttaac aaaatattaa cgcttacaat ttac
//
