LOCUS       sequence-210733-        6104 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of OPN splice variant b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6104)
  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 6104)
  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-OPNb
FEATURES             Location/Qualifiers
     source          1..6104
                     /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(1851..1868)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1857..2081
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2213..2801)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2293..2312)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3130..3177
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3202..3221
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3409..4203)
                     /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(3409..4203)
                     /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(3520..3539)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4130..4149
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4238..4595)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4252..4387
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4404..4595
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4552..4572
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(4655..5446,5447..5515))
                     /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     5278..5297
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5516..5620)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5647..6102
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5734..5753)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5944..5965
                     /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 ctagtccagt
      721 gtgatggata tctgcagaat tcggcttcaa acgccgacca aggaaaactc actaccatga
      781 gaattgcagt gatttgcttt tgcctcctag gcatcacctg tgccatacca gttaaacagg
      841 ctgattctgg aagttctgag gaaaagcagc tttacaacaa atacccagat gctgtggcca
      901 catggctaaa ccctgaccca tctcagaagc agaatctcct agccccacag acccttccaa
      961 gtaagtccaa cgaaagccat gaccacatgg atgatatgga tgatgaagat gatgacgacc
     1021 atgtggacag ccaggactcc attgactcga acgactctga tgatgtagat gacactgatg
     1081 attctcacca gtctgatgag tctcaccatt ctgatgaatc tgatgaactg gtcactgatt
     1141 ttcccacgga cctgccagca accgaagttt tcactccagt tgtccccaca gtagacacat
     1201 acgatggccg aggtgatagt gtagtttatg gactgaggtc aaaatctaag aagtttcgca
     1261 gacctgacat ccagtaccct gatgctacag acgaggacat cacctcacac atggaaagcg
     1321 aggagttgaa tggtgcatac aaggccatcc ccgttgccca ggacctgaac gcgccttctg
     1381 attgggacag ccgtgggaag gacagttatg aaacgagtca gctggatgac cagagtgctg
     1441 aaacccacag ccacaagcag tccagattat ataagcggaa agccaatgat gagagcaatg
     1501 agcattccga tgtgattgat agtcaggaac tttccaaagt cagccgtgaa ttccacagcc
     1561 atgaatttca cagccatgaa gatatgctgg ttgtagaccc caaaagtaag gaagaagata
     1621 aacacctgaa atttcgtatt tctcatgaat tagatagtgc atcttctgag gtcaattaaa
     1681 aggagaaaaa atacaatttc tcactttgca tttagtcaaa agaaaaaatg ctttatagca
     1741 aaatgaaaga gaacatgaaa tgcttctttc tcagtttatt ggttgaagcc gaattccagc
     1801 acagtggcgg ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt
     1861 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     1921 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     1981 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     2041 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     2101 cagtggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2161 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2221 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2281 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2341 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2401 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2461 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2521 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2581 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2641 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2701 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2761 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2821 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     2881 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     2941 aaagtatata tgagtaacct gaggctatgg cagggcctgc cgccccgacg ttggctgcga
     3001 gccctgggcc ttcacccgaa cttggggggt ggggtgggga aaaggaagaa acgcgggcgt
     3061 attggcccca atggggtctc ggtggggtat cgacagagtg ccagccctgg gaccgaaccc
     3121 cgcgtttatg aacaaacgac ccaacaccgt gcgttttatt ctgtcttttt attgccgtca
     3181 tagcgcgggt tccttccggt attgtctcct tccgtgtttc agttagcctc cccctagggt
     3241 gggcgaagaa ctccagcatg agatccccgc gctggaggat catccagccg gcgtcccgga
     3301 aaacgattcc gaagcccaac ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg
     3361 gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc agaagaactc
     3421 gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac
     3481 gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc
     3541 tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg
     3601 gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc
     3661 gccgtcgggc atgctcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg
     3721 ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc
     3781 gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg
     3841 ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag
     3901 atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc
     3961 gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc
     4021 ttgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg
     4081 cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata
     4141 gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat
     4201 catgcgaaac gatcctcatc ctgtctcttg atcgatcttt gcaaaagcct aggcctccaa
     4261 aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat
     4321 aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg
     4381 gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt
     4441 gcatacttct gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga
     4501 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact
     4561 gacacacatt ccacagctgg ttctttccgc ctcaggactc ttcctttttc aataaatcaa
     4621 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4681 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4741 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4801 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4861 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4921 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4981 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5041 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5101 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5161 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5221 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5281 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5341 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5401 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5461 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5521 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5581 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5641 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     5701 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     5761 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     5821 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     5881 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     5941 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     6001 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     6061 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttac
//
