LOCUS       sequence-207709-        6073 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 6073)
  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 6073)
  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 PSM3
FEATURES             Location/Qualifiers
     source          1..6073
                     /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(1811..1828)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1817..2041
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2173..2761)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2253..2272)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3090..3137
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3162..3181
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3369..4163)
                     /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(3369..4163)
                     /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(3480..3499)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4090..4109
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4198..4555)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4212..4347
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4364..4555
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4512..4532
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(4615..5406,5407..5475))
                     /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     5238..5257
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5476..5580)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5607..6062
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5694..5713)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5904..5925
                     /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 aaaagcagaa tgctgtggcc gctgaagaag ccaatgactt taaacaagag
      901 acccttccaa gtaagtccaa cgaaagccat gaccacatgg atgatatgga tgatgaagat
      961 gatgacgacc atgtggacag ccaggactcc attgactcga acgactctga tgatgtagat
     1021 gacactgatg attctcacca gtctgatgag tctcaccatt ctgatgaatc tgatgaactg
     1081 gtcactgatt ttcccacgga cctgccagca accgaagttt tcactccagt tgtccccaca
     1141 gtagacacat atgatggccg aggtgatagt gtggtttatg gactgaggtc aaaatctaag
     1201 aagtttcgca gacctgacat ccagtaccct gatgctacag acgaggacat cacctcacac
     1261 atggaaagcg aggagttgaa tggtgcatac aaggccatcc ccgttgccca ggacctgaac
     1321 gcgccttctg attgggacag ccgtgggaag gacagttatg aaacgagtca gctggatgac
     1381 cagagtgctg aaacccacag ccacaagcag tccagattat ataagcggaa agctaatgat
     1441 gagagcaatg agcattccga tgtgattgat agtcaggaac tttccaaagt cagccgtgaa
     1501 ttccacagcc atgaatttca cagccatgaa gatatgctgg ttgtagaccc caaaagtaag
     1561 gaagaagata aacacctgaa atttcgtatt tctcatgaat tagatagtgc atcttctgag
     1621 gtcaattaaa aggagaaaaa atacaatttc tcactttgca tttagtcaaa agaaaaaatg
     1681 ctttatagca aaatgaaaga gaacatgaaa tgcttctttc tcagtttatt ggttgaagcc
     1741 gaattctgca gatatccatc acactggcgg ccgctcgagt ctagagggcc cgtttaaacc
     1801 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     1861 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     1921 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     1981 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     2041 gcttctgagg cggaaagaac cagtggcggt aatacggtta tccacagaat caggggataa
     2101 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2161 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2221 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2281 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2341 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2401 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2461 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2521 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2581 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     2641 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2701 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2761 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     2821 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     2881 atgaagtttt aaatcaatct aaagtatata tgagtaacct gaggctatgg cagggcctgc
     2941 cgccccgacg ttggctgcga gccctgggcc ttcacccgaa cttggggggt ggggtgggga
     3001 aaaggaagaa acgcgggcgt attggcccca atggggtctc ggtggggtat cgacagagtg
     3061 ccagccctgg gaccgaaccc cgcgtttatg aacaaacgac ccaacaccgt gcgttttatt
     3121 ctgtcttttt attgccgtca tagcgcgggt tccttccggt attgtctcct tccgtgtttc
     3181 agttagcctc cccctagggt gggcgaagaa ctccagcatg agatccccgc gctggaggat
     3241 catccagccg gcgtcccgga aaacgattcc gaagcccaac ctttcataga aggcggcggt
     3301 ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag
     3361 agtcccgctc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga
     3421 gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca
     3481 atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag
     3541 tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca
     3601 tgggtcacga cgagatcctc gccgtcgggc atgctcgcct tgagcctggc gaacagttcg
     3661 gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc
     3721 atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc
     3781 ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga
     3841 gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt
     3901 cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac
     3961 gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac cggacaggtc ggtcttgaca
     4021 aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt
     4081 gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg
     4141 tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcgatcttt
     4201 gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga
     4261 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
     4321 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
     4381 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     4441 cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     4501 ggactttcca caccctaact gacacacatt ccacagctgg ttctttccgc ctcaggactc
     4561 ttcctttttc aataaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4621 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4681 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4741 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4801 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4861 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4921 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4981 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5041 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5101 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5161 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5221 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5281 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5341 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5401 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5461 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5521 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5581 cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg
     5641 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     5701 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     5761 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     5821 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     5881 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     5941 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     6001 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     6061 ttacgcgcgc gtt
//
