LOCUS       sequence-207221-        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 M1
FEATURES             Location/Qualifiers
     source          1..6073
                     /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(1820..1837)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1826..2050
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2182..2770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2262..2281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3099..3146
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3171..3190
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3378..4172)
                     /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(3378..4172)
                     /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(3489..3508)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4099..4118
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4207..4564)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4221..4356
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4373..4564
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4521..4541
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(4624..5415,5416..5484))
                     /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     5247..5266
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5485..5589)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5616..6071
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5703..5722)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5913..5934
                     /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 gttctgcggc agcggcgaat gctgtgtcct ctgaagaaac caatgacttt
      901 aaacaagaga cccttccaag taagtccaac gaaagccatg accacatgga tgatatggat
      961 gatgaagatg atgacgacca tgtggacagc caggactcca ttgactcgaa cgactctgat
     1021 gatgtagatg acactgatga ttctcaccag tctgatgagt ctcaccattc tgatgaatct
     1081 gatgaactgg tcactgattt tcccacggac ctgccagcaa ccgaagtttt cactccagtt
     1141 gtccccacag tagacacata tgatggccga ggtgatagtg tggtttatgg actgaggtca
     1201 aaatctaaga agtttcgcag acctgacatc cagtaccctg atgctacaga cgaggacatc
     1261 acctcacaca tggaaagcga ggagttgaat ggtgcataca aggccatccc cgttgcccag
     1321 gacctgaacg cgccttctga ttgggacagc cgtgggaagg acagttatga aacgagtcag
     1381 ctggatgacc agagtgctga aacccacagc cacaagcagt ccagattata taagcggaaa
     1441 gctaatgatg agagcaatga gcattccgat gtgattgata gtcaggaact ttccaaagtc
     1501 agccgtgaat tccacagcca tgaatttcac agccatgaag atatgctggt tgtagacccc
     1561 aaaagtaagg aagaagataa acacctgaaa tttcgtattt ctcatgaatt agatagtgca
     1621 tcttctgagg tcaattaaaa ggagaaaaaa tacaatttct cactttgcat ttagtcaaaa
     1681 gaaaaaatgc tttatagcaa aatgaaagag aacatgaaat gcttctttct cagtttattg
     1741 gttgaagccg aattctgcag atatccatca cactggcggc cgctcgagtc tagagggccc
     1801 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     1861 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     1921 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     1981 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     2041 ggctctatgg cttctgaggc ggaaagaacc agtggcggta atacggttat ccacagaatc
     2101 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2161 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2221 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2281 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2341 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     2401 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2461 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     2521 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2581 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     2641 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     2701 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2761 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     2821 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     2881 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaacctg aggctatggc
     2941 agggcctgcc gccccgacgt tggctgcgag ccctgggcct tcacccgaac ttggggggtg
     3001 gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg gtggggtatc
     3061 gacagagtgc cagccctggg accgaacccc gcgtttatga acaaacgacc caacaccgtg
     3121 cgttttattc tgtcttttta ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt
     3181 ccgtgtttca gttagcctcc ccctagggtg ggcgaagaac tccagcatga gatccccgcg
     3241 ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa
     3301 ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc
     3361 gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc
     3421 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc
     3481 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc
     3541 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag
     3601 gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg
     3661 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga
     3721 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg
     3781 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc
     3841 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc
     3901 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg
     3961 gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg
     4021 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag
     4081 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga
     4141 gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga
     4201 tcgatctttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
     4261 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
     4321 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
     4381 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     4441 ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     4501 ggagcctggg gactttccac accctaactg acacacattc cacagctggt tctttccgcc
     4561 tcaggactct tcctttttca ataaatcaat ctaaagtata tatgagtaaa cttggtctga
     4621 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4681 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4741 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4801 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4861 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4921 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4981 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5041 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5101 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5161 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5221 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5281 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5341 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5401 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5461 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5521 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5581 ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag
     5641 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     5701 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     5761 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     5821 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     5881 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     5941 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     6001 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     6061 gcttacaatt tac
//
