LOCUS       sequence-147479-        6570 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 6570)
  AUTHORS   Shukla AK, Kim J, Ahn S, Xiao K, Shenoy SK, Liedtke W, Lefkowitz RJ
  TITLE     Arresting a transient receptor potential (TRP) channel: 
            beta-arrestin 1 mediates ubiquitination and functional 
            down-regulation of TRPV4.
  JOURNAL   J Biol Chem. 2010 Sep 24;285(39):30115-25. doi: 
            10.1074/jbc.M110.141549. Epub 2010 Jul 22.
  PUBMED    20650893
REFERENCE   2  (bases 1 to 6570)
  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: 
            "10.1074/jbc.M110.141549"; journalName: "J Biol Chem"; date: 
            "2010-09-24- 24"; volume: "285"; issue: "39"; pages: "30115-25"
COMMENT     Sequence Label: pcDNA3.0 FLAG TRPV4
FEATURES             Location/Qualifiers
     source          1..6570
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(63..80)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(234..822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(314..333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(993..1784,1785..1853))
                     /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     1616..1635
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1854..1958)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1984..2439)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2121..2142)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2333..2352
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3318..3336
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3318..3335
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             6022..6045
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    6112..6246
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6136..6155)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6190..6209
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6363..6383)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6363..6381)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6402..6418)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6402..6418)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6415..6437)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6426..6442
                     /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(6450..6456,6457..6474,6475..6480))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6495..6516
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
       61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     1861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1981 cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     2041 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     2101 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     2161 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     2221 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     2281 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     2341 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     2401 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc
     2461 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag tcgatcgacc
     2521 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
     2581 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
     2641 gccaattcaa tatggctatt ggccaggttc aatactatgt attggcccta tgccatatag
     2701 tattccatat atgggttttc ctattgacgt agatagcccc tcccaatggg cggtcccata
     2761 taccatatat ggggcttcct aataccgccc atagccactc ccccattgac gtcaatggtc
     2821 tctatatatg gtctttccta ttgacgtcat atgggcggtc ctattgacgt atatggcgcc
     2881 tcccccattg acgtcaatta cggtaaatgg cccgcctggc tcaatgccca ttgacgtcaa
     2941 taggaccacc caccattgac gtcaatggga tggctcattg cccattcata tccgttctca
     3001 cgccccctat tgacgtcaat gacggtaaat ggcccacttg gcagtacatc aatatctatt
     3061 aatagtaact tggcaagtac attactattg gaagtacgcc agggtacatt ggcagtactc
     3121 ccattgacgt caatggcggt aaatggcccg cgatggctgc caagtacatc cccattgacg
     3181 tcaatgggga ggggcaatga cgcaaatggg cgttccattg acgtaaatgg gcggtaggcg
     3241 tgcctaatgg gaggtctata taagcaatgc tcgtttaggg aaccgccatt ctgcctgggg
     3301 acgtcggagc aagcttgatt taggtgacac tatagaatac aagctacttg ttctttttgc
     3361 aggatcaagc aggccgagaa gtccaaacag atctgctcag ggtccagtat ggcagatcct
     3421 ggtgatggcc cccgtgcagc gcctggggat gtggctgagc cccctggaga cgagagtggc
     3481 acttctggtg gggaggcctt ccccctctct tccctggcca acctgtttga gggagaggaa
     3541 ggctcctctt ctctttcacc agtggatgct agccgccctg ctggccccgg ggatggacgt
     3601 ccaaacctgc gtatgaagtt ccagggcgct ttccgcaagg gggttcccaa ccccattgac
     3661 ctgctggagt ccaccctgta tgagtcctca gtagtgcctg ggcccaagaa agcgcccatg
     3721 gattcgttgt tcgactatgg cacttaccgg caccacccca gtgacaacaa gagatggagg
     3781 aggaaggtcg tagagaagca gccacagagc cccaaagctc ccgcccccca gccacccccc
     3841 atcctcaaag tcttcaaccg gcccatcctc tttgacatcg tgtcccgggg ctccactgcc
     3901 gacctggacg gactgctctc ctacttgctg acccacaaga agcgcctgac tgatgaggag
     3961 ttccgggaac catccacagg gaagacctgc ctgcccaagg cacttctgaa cttaagcaat
     4021 ggccgaaacg acaccatccc agtgttgctg gacattgcgg aacgcacggg caacatgcgg
     4081 gagttcatca actcgccctt cagagacatc tactaccgag ggcagacggc actgcacatc
     4141 gccattgaac ggcgctgcaa gcattacgtg gagctcctgg tggcccaggg agccgatgtg
     4201 cacgcgcagg cccgagggcg gttcttccag cccaaggatg agggtggcta cttctacttt
     4261 ggggagctgc ccttgtcctt ggcagcctgc accaaccagc cgcacatcgt caactacctg
     4321 acagagaacc ctcacaagaa agccgatatg aggcgacagg actccagagg caacacggtg
     4381 ctccacgcgc tggtggccat cgctgacaac acccgagaga acaccaagtt tgtcaccaag
     4441 atgtatgacc tgttgcttct caagtgctcc cgcctcttcc cagacagcaa cctggagact
     4501 gtgcttaaca atgacggtct ttcgcccctc atgatggctg ccaagactgg caagatcggg
     4561 gtctttcagc acatcatccg acgggaggtg acagatgagg acacacggca cctgtctcgc
     4621 aagttcaagg actgggccta cgggcctgtg tattcttctc tctacgacct ctcctccctg
     4681 gatacgtgcg gggaggaagt gtccgtgctg gagatcctgg tttacaacag caagatcgag
     4741 aaccgccatg agatgctggc tgtggagccc attaacgaac tgctgaggga caagtggcgt
     4801 aagttcgggg ccgtgtcctt ctacatcaac gttgtctcct atctgtgtgc catggtcatc
     4861 ttcaccctca cagcctacta tcagccactg gagggcacgc caccctaccc ttaccgtacc
     4921 acggtggact acctgaggct ggctggtgag gtcatcacgc tcctcacagg agtcctgttc
     4981 ttctttacca gtatcaaaga cttgttcatg aagaaatgcc ctggagtgaa ttctctcttc
     5041 gtcgatggct ccttccagtt gctctacttc atctactcag tgctggtggt tgtgtctgcg
     5101 gcgctctacc tggcagggat cgaggcctat ctggctgtga tggtctttgc cctggtcctg
     5161 ggctggatga atgcccttta cttcacccgt gggctgaagc tgacagggac ctacagcatc
     5221 atgattcaga agatcctctt caaagatctc ttccgctttc tgctggtcta cctgcttttt
     5281 atgattggct atgcctcagc tctggtcacc ctcctgaatc cgtgcaccaa catgaaggtc
     5341 tgtaacgagg accagagcaa ctgcacggtg ccctcatacc ccgcgtgccg ggacagcgag
     5401 accttcagcg ccttcctact ggacctcttc aagctcacca tcggcatggg cgacctggag
     5461 atgctgagca gcgctaagta ccccgtggtc ttcattctcc tgctggttac ctacatcatc
     5521 ctcaccttcg tgctcctgct gaacatgctc atcgccctca tgggtgagac cgtgggccag
     5581 gtgtccaagg agagcaagca catctggaag ctgcagtggg ccaccaccat cctggacatc
     5641 gagcgctcct tccctgtgtt cctgaggaag gccttccgct ccggagagat ggtgacagtg
     5701 ggcaagagct cggatggcac tccagaccgc aggtggtgct tcagggtgga cgaggtgaac
     5761 tggtctcact ggaaccagaa cctgggcatc attaacgagg accccggcaa gagcgagatc
     5821 taccagtact atggcttctc ccataccatg gggcgcctcc gcagggatcg ctggtcctca
     5881 gtggtgcccc gcgtggtgga gctgaacaag aactcaggca cagatgaagt ggtggtcccc
     5941 ctggataacc tagggaaccc caactgtgac ggccaccagc aaggttatgc tcccaagtgg
     6001 agggcggagg acgcaccact ggattacaag gacgatgacg ataaggtgac tataaagacg
     6061 aagacgataa atgactcgag cctctagaac tatagtgagt cgtattacgt agatccagac
     6121 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     6181 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     6241 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     6301 gttttttaat tcgcggcgcg ccgcggcgcc aatgcattgg gcccggtacc cagcttttgt
     6361 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
     6421 tgaaattgtt atccgctcac aattccacac aacatacgag ccgggagcat aaagtgtaaa
     6481 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6541 ttccagtcgg gaaacctgtc gtgccagctg
//
