LOCUS       sequence-177967-        6374 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Knockdown human STIM1 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6374)
  AUTHORS   Sun J, Lu F, He H, Shen J, Messina J, Mathew R, Wang D, Sarnaik AA, 
            Chang WC, Kim M, Cheng H, Yang S
  TITLE     STIM1- and Orai1-mediated Ca(2+) oscillation orchestrates 
            invadopodium formation and melanoma invasion.
  JOURNAL   J Cell Biol. 2014 Nov 24;207(4):535-48. doi: 10.1083/jcb.201407082. 
            Epub 2014 Nov 17.
  PUBMED    25404747
REFERENCE   2  (bases 1 to 6374)
  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: 
            "10.1083/jcb.201407082"; journalName: "J Cell Biol"; date: 
            "2014-11-24- 24"; volume: "207"; issue: "4"; pages: "535-48"
COMMENT     Sequence Label: pSUPER-retro-puro-shSTIM1
FEATURES             Location/Qualifiers
     source          1..6374
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..215
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     136..155
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     primer_bind     complement(286..302)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(287..303)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             complement(321..737)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(337..353)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(369..391)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    804..1145
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             1209..1725
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(1925..1944)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(2072..2094)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2288..2307)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2407..2429
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2467..2485)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2553..2657
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2658..2726,2727..3518)
                     /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     complement(2876..2895)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3689..4277
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4178..4197
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4431..4448
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4565..4586
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4601..4606,4607..4624,4625..4631)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4639..4655
                     /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             complement(5241..5840)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5324..5344)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     5821..5840
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(5865..6364)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5913..5932)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     6306..6329
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 gaacgctgac gtcatcaacc cgctccaagg aatcgcgggc ccagtgtcac taggcgggaa
       61 cacccagcgc gcgtgcgccc tggcaggaag atggctgtga gggacagggg agtggcgccc
      121 tgcaatattt gcatgtcgct atgtgttctg ggaaatcacc ataaacgtga aatgtctttg
      181 gatttgggaa tcttataagt tctgtatgag accacagatc cccagaagga gctagaatct
      241 cacttcaaga gagtgagatt ctagctcctt cttttttaag cttatcgata ccgtcgacct
      301 cgagagatcg atctaattcc ggcgcctaga gaaggagtga gggctggata aagggaggat
      361 cgaggcgggg tcgaacgagg aggttcaagg gggagagacg gggcggatgg aggaagagga
      421 ggcggaggct tagggtgtac aaagggcttg acccagggag gggggtcaaa agccaaggct
      481 tcccaggtca cgatgtaggg gacctggtct gggtgtccat gcgggccagg tgaaaagacc
      541 ttgatcttaa cctgggtgat gaggtctcgg ttaaaggtgc cgtctcgcgg ccatccgacg
      601 ttaaaggttg gccattctgc agagcagaag gtaacccaac gtctcttctt gacatctacc
      661 gactggttgt gagcgatccg ctcgacatct ttccagtgac ctaaggtcaa acttaaggga
      721 gtggtaacag tctggcccta attttcagac aaatacagaa acacagtcag acagagacaa
      781 cacagaacga tgctgcagcg ctgcagcaga caagacgcgc ggcttcggtt ccaaaccgaa
      841 agcaaaaatt cagacggagg cgggaactgt tttaggttct cgtctcctac cagaaccaca
      901 tatcctgacg gggtcggatt ccacatcgac tcccttcctc aggtcgggcc acaaaaacgg
      961 cccccaaagt ccctgggacg tctcccaggg ttgcggccgg gtgttcagaa ctcgtcagtt
     1021 ccaccacggg tccgccagat acagagctag ttagctaact agtacagacg caggcgcaaa
     1081 cattagatgc cggcacaaac acgcacaaag acagagacag acacgaaacg accgctggcc
     1141 agcttacctc ccggcggggg ggtcggtggt ccctgggcag gggtctccaa atctcggtgg
     1201 aacctccaaa tgaaagaccc ccgaggtggg cagtcaatca atctgaggag accctcccaa
     1261 ggatcagcga gtccacgatt cggatgcaaa cagcaagagg ctttattggg aatacgggta
     1321 cccgggcgac gcagtctatc ggaggactgg cgcgccgagt gaggggttgt gggctctttt
     1381 attgagctcg gggagcagaa gcgcgcgaac agaagcgaga agcgaactga ttggttagtt
     1441 caaataaggc acagggtcat ttcaggtcct tggggcaccc tggaaacatc tgatggttct
     1501 ctagaaactg ctgagggcgg gaccgcatct ggggaccatc tgttcttggc cctgagccgg
     1561 ggcaggaact gcttaccaca gatatcctgt ttggcccata ttctgctgtc tctctgttcc
     1621 taaccttgat ctgaacttct ctattctcag ttatgtattt tccatgcctt gcaaaatggc
     1681 gttacttaag ctagcttgcc aaacctacag gtggggtctt tcattccccc ctttttctgg
     1741 agactaaata aaatctttta ttttatctat ggctcgtact ctataggctt cagctggtga
     1801 tattgttgag tcaaaactag agcctggacc actgatatcc tgtctttaac aaattggact
     1861 aatcgcctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg tgcggttgct
     1921 ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt cgggctcatg
     1981 agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact gttgggcgcc
     2041 atctccttgc atgcaccatt ccttgcgccg tcgttttaca acgtcgtgac tgggaaaacc
     2101 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     2161 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
     2221 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca
     2281 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     2341 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     2401 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     2461 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     2521 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2581 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2641 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2701 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2761 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2821 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     2881 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     2941 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     3001 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     3061 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     3121 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     3181 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     3241 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     3301 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3361 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3421 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3481 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3541 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3601 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3661 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3721 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3781 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     3841 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3901 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3961 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4021 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4081 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4141 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4201 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4261 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4321 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4381 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     4441 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     4501 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     4561 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     4621 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca caattctccc cagctctttc
     4681 tgcgctgtcc ataatgaact atttcaggag tattggagta tgtggattca aaacagtggc
     4741 taattgagtc aggattactt aatattcgac tcctaccctc agaatgttgt tctccaactt
     4801 caagaaactg ttacccatga aagacccccg ctgacgggta gtcaatcact cagaggagac
     4861 cctcccaagg aacagcgaga ccacaagtcg gatgcaactg caagagggtt tattggatac
     4921 acgggtaccc gggcgacgca gtctatcgga ggactggcgc gccgagtgag gggttgtggg
     4981 ctcttttatt gagctcgggg agcagaagcg cgcgaacaga agcgagaagc gaactgattg
     5041 gttagttcaa ataaggcaca gggtcatttc aggtccttgg ggcaccctgg aaacatctga
     5101 tggttctcta gcttgccaaa cctacaggtg gggtctttca ttcccccctt tttctggaga
     5161 ctaaataaaa tcttttattt tatcgatgca tggggtcgtg cgctcctttc ggtcgggcgc
     5221 tgcgggtcgt ggggcgggcg tcaggcaccg ggcttgcggg tcatgcacca ggtgcgcggt
     5281 ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga gccgctcgta gaaggggagg
     5341 ttgcggggcg cggaggtctc caggaaggcg ggcaccccgg cgcgctcggc cgcctccact
     5401 ccggggagca cgacggcgct gcccagaccc ttgccctggt ggtcgggcga gacgccgacg
     5461 gtggccagga accacgcggg ctccttgggc cggtgcggcg ccaggaggcc ttccatctgt
     5521 tgctgcgcgg ccagccggga accgctcaac tcggccatgc gcgggccgat ctcggcgaac
     5581 accgcccccg cttcgacgct ctccggcgtg gtccagaccg ccaccgcggc gccgtcgtcc
     5641 gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga ggaagagttc ttgcagctcg
     5701 gtgacccgct cgatgtggcg gtccggatcg acggtgtggc gcgtggcggg gtagtcggcg
     5761 aacgcggcgg cgagggtgcg tacggccctg gggacgtcgt cgcgggtggc gaggcgcacc
     5821 gtgggcttgt actcggtcat ggtaagctag cttgggctgc aggtcgaaag gcccggagat
     5881 gaggaagagg agaacagcgc ggcagacgtg cgcttttgaa gcgtgcagaa tgccgggcct
     5941 ccggaggacc ttcgggcgcc cgccccgccc ctgagcccgc ccctgagccc gcccccggac
     6001 ccaccccttc ccagcctctg agcccagaaa gcgaaggagc aaagctgcta ttggccgctg
     6061 ccccaaaggc ctacccgctt ccattgctca gcggtgctgt ccatctgcac gagactagtg
     6121 agacgtgcta cttccatttg tcacgtcctg cacgacgcga gctgcggggc gggggggaac
     6181 ttcctgacta ggggaggagt agaaggtggc gcgaaggggc caccaaagaa cggagccggt
     6241 tggcgcctac cggtggatgt ggaatgtgtg cgaggccaga ggccacttgt gtagcgccaa
     6301 gtgcccagcg gggctgctaa agcgcatgct ccagactgcc ttgggaaaag cgcctcccct
     6361 acccggtaga attc
//
