LOCUS       sequence-209939-        5859 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of OPN splice variant c.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5859)
  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 5859)
  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: pGEX-6P1-OPNc-delta S
FEATURES             Location/Qualifiers
     source          1..5859
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(183..188,189..204,205..211)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    219..235
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     869..891
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             918..941
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(1909..1931)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2076..2094)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2162..2266
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2267..2335,2336..3127)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2485..2504)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3298..3886
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3787..3806
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4040..4057
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4130..4207
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4208..5290
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4227..4246)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5339..5344,5345..5362,5363..5369)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5377..5393
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5401..5417
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5401..5417
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5413..5859,1..81)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5433..5450)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5433..5449)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5442..5464)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 agcttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg gaagttctgt tccaggggcc cctgggatcc ataccagtta
      961 aacaggctga ttctggaagt tctgaggaaa agcagaatgc tgtgtcctct gaagaaacca
     1021 atgactttaa acaagagacc cttccaagta agtccaacga aagccatgac cacatggatg
     1081 atatggatga tgaagatgat gacgaccatg tggacagcca ggactccatt gactcgaacg
     1141 actctgatga tgtagatgac actgatgatt ctcaccagtc tgatgagtct caccattctg
     1201 atgaatctga tgaactggtc actgattttc ccacggacct gccagcaacc gaagttttca
     1261 ctccagttgt ccccacagta gacacatatg atggccgagg tgatagtgtg gtttatggac
     1321 tgaggtcaaa atctaagaag tttcgcagac ctgacatcca gtaccctgat gctacagacg
     1381 aggacatcac ctcacacatg gaaagcgagg agttgaatgg tgcatacaag gccatccccg
     1441 ttgcccagga cctgaacgcg ccttctgatt gggacagccg tgggaaggac agttatgaaa
     1501 cgagtcagct ggatgaccag agtgctgaaa cccacagcca caagcagtcc agattatata
     1561 agcggaaagc taatgatgag agcaatgagc attccgatgt gattgatagt caggaacttt
     1621 ccaaagtcag ccgtgaattc cacagccatg aatttcacag ccatgaagat atgctggttg
     1681 tagaccccaa aagtaaggaa gaagataaac acctgaaatt tcgtatttct catgaattag
     1741 atagtgcatc ttctgaggtc aattaaaagg agaaaaaata caatttctca ctttgcattt
     1801 agtcaaaaga aaaaatgctt tatagcaaaa tgaaagagaa catctcgagc ggccgcatcg
     1861 tgactgactg acgatctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     1921 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     1981 tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag
     2041 cgatagcgga gtgtataatt cttgaagacg aaagggcctc gtgatacgcc tatttttata
     2101 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     2161 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2221 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     2281 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     2341 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     2401 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     2461 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     2521 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     2581 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     2641 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     2701 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     2761 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc
     2821 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     2881 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     2941 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     3001 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     3061 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     3121 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     3181 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     3241 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3301 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3361 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3421 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     3481 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3541 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3601 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3661 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3721 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3781 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3841 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3901 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     3961 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     4021 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     4081 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc ataaattccg acaccatcga
     4141 atggcgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc aattcagggt
     4201 ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg tctcttatca
     4261 gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc gggaaaaagt
     4321 ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac aactggcggg
     4381 caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg cgccgtcgca
     4441 aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat
     4501 ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg
     4561 cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg ctgtggaagc
     4621 tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac ccatcaacag
     4681 tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg tcgcattggg
     4741 tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc gtctgcgtct
     4801 ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg
     4861 cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt
     4921 tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac
     4981 cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg ataccgaaga
     5041 cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc tgctggggca
     5101 aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct
     5161 gttgcccgtc tcactggtga aaagaaaaac cacgctagcg cccaatacgc aaaccgcctc
     5221 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     5281 cgggcagtga gcgcaacgca attaatgtaa gttagctcac tcattaggca ccccaggctt
     5341 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     5401 caggaaacag ctatgaccat gattacggat tcactggccg tcgttttaca acgtcgtgac
     5461 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     5521 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     5581 ggcgaatggc gctttgcctg gtttccggca ccagaagcgg tgccggaaag ctggctggag
     5641 tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa actggcagat gcacggttac
     5701 gatgcgccca tctacaccaa cgtgacctat cccattacgg tcaatccgcc gtttgttccc
     5761 acggagaatc cgacgggttg ttactcgctc acatttaatg ttgatgaaag ctggctacag
     5821 gaaggccaga cgcgaattat ttttgatggc gttggaatt
//
