LOCUS       sequence-140174-        5188 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5188)
  AUTHORS   Lian N, Wang W, Li L, Elefteriou F, Yang X
  TITLE     Vimentin inhibits ATF4-mediated osteocalcin transcription and 
            osteoblast differentiation.
  JOURNAL   J Biol Chem. 2009 Oct 30;284(44):30518-25. Epub 2009 Sep 2.
  PUBMED    19726676
REFERENCE   2  (bases 1 to 5188)
  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"; journalName: "J Biol 
            Chem."; date: "2009-10-30"; volume: "284(44)"; pages: "30518-25. 
            Epub 2009 Sep 2"
COMMENT     Sequence Label: HA-Vim
FEATURES             Location/Qualifiers
     source          1..5188
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     505..529
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          646..742
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             806..832
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2301..2435
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2325..2344)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2379..2398
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2546..2562)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2546..2562)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2559..2581)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2570..2586
                     /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)."
     promoter        complement(join(2594..2600,2601..2618,2619..2624))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2639..2660
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2777..2794)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2948..3536)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3028..3047)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3707..4498,4499..4567))
                     /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     4330..4349
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4568..4672)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4740..4758
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4796..4818)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4918..4937
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5131..5153
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5145..5162
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5146..5162
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      541 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccggactct
      601 agaggatccg gtactagagg aactgaaaaa ccagaaagtt aactggtaag tttagtcttt
      661 ttgtctttta tttcaggtcc cggatccggt ggtggtgcaa atcaaagaac tgctcctcag
      721 tggatgttgc ctttacttct aggcctgtac ggaagtgtta cttctgctct aaaagctgcg
      781 gaattgtacc cgcgggccca ccatgtaccc atacgatgtt ccagattacg ctcttatggc
      841 catggaggcc cgaattctgt ctaccaggtc tgtgtcctcg tcctcctacc gcaggatgtt
      901 cggtggctcc ggcacatcga gccggcccag ctccaaccgg agctatgtga ccacgtccac
      961 acgcacctac agtctgggca gcgcactgcg ccccagcact agccgcagcc tctattcctc
     1021 atcccccggt ggcgcctatg tgacccggtc ctcggcagtg cgcctgcgga gcagcgtgcc
     1081 gggcgtgcgg ctgcttcaag actcggtgga cttctcgctg gccgacgcca tcaacactga
     1141 gttcaagaac acccgcacca acgagaaggt agaactgcag gagctgaatg accgctttgc
     1201 caactacatc gacaaggtgc gcttcctcga gcagcagaac aaaatcctgc tggctgagct
     1261 cgagcagctc aagggccagg gcaagtcgcg cctgggcgac ctgtacgagg aggagatgcg
     1321 ggagctgcgc cggcaggtgg atcagctcac caacgacaag gcccgtgtcg aggtggagcg
     1381 ggacaacctg gccgaggaca tcatgcggct gcgagagaaa ttgcaggagg agatgctcca
     1441 gagagaggaa gccgaaagca ccctgcagtc attcagacag gatgttgaca atgcttctct
     1501 ggcacgtctt gaccttgaac ggaaagtgga atccttgcag gaagaaattg cctttttgaa
     1561 gaaactgcac gatgaagaga tccaggagct gcaggcccag attcaggaac agcatgtcca
     1621 gatcgatgtg gacgtttcca agcctgacct cactgctgcc ctgcgtgatg tgcgccagca
     1681 gtatgaaagc gtggctgcca agaacctcca ggaggccgag gaatggtaca agtccaagtt
     1741 tgctgacctc tctgaggctg ccaaccggaa caacgatgcc ctgcgccagg ccaagcagga
     1801 gtcaaacgag taccggagac aggtgcagtc actcacctgt gaagtggatg cccttaaagg
     1861 cactaacgag tccctggagc gccagatgcg tgagatggaa gagaattttg cccttgaagc
     1921 tgctaactac caggacacta ttggccgcct gcaggatgag atccaaaaca tgaaggaaga
     1981 gatggctcgt caccttcgtg aataccaaga tctgctcaat gttaagatgg ccctggacat
     2041 tgagatcgcc acctacagga agctgctgga aggcgaggag agcaggattt ctctgcctct
     2101 gccaaccttt tcttccctga acctgagaga aactaacctg gagtcacttc ctctggttga
     2161 cacccactca aaaagaacac tcctgattaa gacggttgag accagagatg gacaggtgat
     2221 caatgagact tctcagcatc acgatgacct tgaataaaaa ttgcacacac ttggatctct
     2281 cgaggtaccg cggccgcggg gatccagaca tgataagata cattgatgag tttggacaaa
     2341 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     2401 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     2461 tgtttcaggt tcagggggag gtgtgggagg ttttttcgga tcctctagag tcgatctgca
     2521 ggcatgctag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     2581 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     2641 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     2701 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     2761 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2821 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2881 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2941 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3001 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3061 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3121 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3181 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3241 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3301 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3361 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3421 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3481 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3541 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3601 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3661 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3721 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3781 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3841 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3901 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     3961 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4021 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4081 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4141 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4201 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4261 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4321 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4381 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4441 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4501 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4561 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     4621 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     4681 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     4741 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     4801 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     4861 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     4921 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     4981 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
     5041 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     5101 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     5161 gtgagttcga gcttgcatgc ctgcaggt
//
