LOCUS       sequence-59317-e        6670 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 6670)
  AUTHORS   Talbot JJ, Shillingford JM, Vasanth S, Doerr N, Mukherjee S, Kinter 
            MT, Watnick T, Weimbs T
  TITLE     Polycystin-1 regulates STAT activity by a dual mechanism.
  JOURNAL   Proc Natl Acad Sci U S A. 2011 May 10;108(19):7985-90. Epub 2011 Apr
            25.
  PUBMED    21518865
REFERENCE   2  (bases 1 to 6670)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2011-05-10"; volume: "108(19)"; pages: 
            "7985-90. Epub 2011 Apr 25"
COMMENT     Sequence Label: FLM-PC1
FEATURES             Location/Qualifiers
     source          1..6670
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    41..59
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    62..80
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     90..114
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1774..1803
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1813..1842
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1858..1875
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1898..1915)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1904..2128
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2174..2602
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2261..2280)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2471..2492
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2611..2631)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2616..2946
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2797..2932
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2859..2878
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2994..3041
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3060..3434
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    3564..3685
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3601..3620)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3655..3674
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3734..3750)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3734..3750)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3747..3769)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3758..3774
                     /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(3782..3788,3789..3806,3807..3812))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3827..3848
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3965..3982)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4136..4724)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4216..4235)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4895..5686,5687..5755))
                     /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     5518..5537
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5756..5860)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5935..5954)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6126..6505
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(6506..6670,1..39)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     join(6660..6670,1..10)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 ggtaggcgtg tacggtggga ggtctatata agcagagctc tccctatcag tgatagagat
       61 ctccctatca gtgatagaga tcgtcgacga gctcgtttag tgaaccgtca gatcgcctgg
      121 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
      181 actctagcgt ttaaacttaa gcttctagag atccctcgac ctcgagatcc attgtgctct
      241 aaagatcttt ggtgacttgt ccactccagt gtggcatcat gtggcagctg ctcctcccaa
      301 ctgctctgct acttctagtt tcagctggca tgcggactga agatctccca aaggctgtgg
      361 tgttcctgga gcctcaatgg tacagcgtgc ttgagaagga cagtgtgact ctgaagtgcc
      421 agggagccta ctcccctgag gacaattcca cacagtggtt tcacaatgag agcctcatct
      481 caagccaggc ctcgagctac ttcattgacg ctgccacagt caacgacagt ggagagtaca
      541 ggtgccagac aaacctctcc accctcagtg acccggtgca gctagaagtc catatcggct
      601 ggctgttgct ccaggcccct cggtgggtgt tcaaggagga agaccctatt cacctgaggt
      661 gtcacagctg gaagaacact gctctgcata aggtcacata tttacagaat ggcaaagaca
      721 ggaagtattt tcatcataat tctgacttcc acattccaaa agccacactc aaagatagcg
      781 gctcctactt ctgcaggggg cttgttggga gtaaaaatgt gtcttcagag actgtgaaca
      841 tcaccatcac tcaaggtttg gcagtgtcaa ccatctcatc attctctcca cctggcgcgg
      901 atccaccaag ggcctctgcc ctccctgccc caccgacagg ctccgccctc cctgacccgc
      961 agacagcctc tgccctccct gacccgccag cagcctctgc cctccctgcg gccctggcgg
     1021 tgatctcctt cctcctcggg ctgggcctgg gggtggcgtg tgtgctggcg aggacgcgtg
     1081 agtcctggca cttgtcaccc ctgctgtgtg tggggctctg ggcactgcgg ctgtggggcg
     1141 ccctacggct gggggctgtt attctccgct ggcgctacca cgccttgcgt ggagagctgt
     1201 accggccggc ctgggagccc caggactacg agatggtgga gttgttcctg cgcaggctgc
     1261 gcctctggat gggcctcagc aaggtcaagg agttccgcca caaagtccgc tttgaaggga
     1321 tggagccgct gccctctcgc tcctccaggg gctccaaggt atccccggat gtgcccccac
     1381 ccagcgctgg ctccgatgcc tcgcacccct ccacctcctc cagccagctg gatgggctga
     1441 gcgtgagcct gggccggctg gggacaaggt gtgagcctga gccctcccgc ctccaagccg
     1501 tgttcgaggc cctgctcacc cagtttgacc gactcaacca ggccacagag gacgtctacc
     1561 agctggagca gcagctgcac agcctgcaag gccgcaggag cagccgggcg cccgccggat
     1621 cttcccgtgg cccatccccg ggcctgcggc cagcactgcc cagccgcctt gcccgggcca
     1681 gtcggggtgt ggacctggcc actggcccca gcaggacacc ccttcgggcc aagaacaagg
     1741 tccaccccag cagcacttag ggcggccgct cgagaacaaa aactcatctc agaagaggat
     1801 ctggggccct tcgaacaaaa actcatctca gaagaggatc tgaatatgca taccggtcat
     1861 catcaccatc accattgagt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     1921 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     1981 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     2041 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     2101 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     2161 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2221 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     2281 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta
     2341 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     2401 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     2461 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     2521 tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt
     2581 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     2641 ccccaggctc cccaggcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2701 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2761 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     2821 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     2881 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     2941 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgttgaca
     3001 attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg aactaaacca
     3061 tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg
     3121 agttctggac cgaccggctc gggttctccc gggacttcgt ggaggacgac ttcgccggtg
     3181 tggtccggga cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca
     3241 acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag tggtcggagg
     3301 tcgtgtccac gaacttccgg gacgcctccg ggccggccat gaccgagatc ggcgagcagc
     3361 cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg
     3421 aggagcagga ctgacacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt
     3481 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     3541 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     3601 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3661 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     3721 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3781 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3841 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3901 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3961 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     4021 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4081 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4141 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4201 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4261 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4321 cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4381 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4441 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4501 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4561 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4621 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4681 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4741 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4801 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4861 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4921 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4981 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     5041 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     5101 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     5161 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     5221 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5281 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5341 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5401 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5461 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5521 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5581 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5641 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5701 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5761 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5821 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5881 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat
     5941 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     6001 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     6061 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
     6121 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     6181 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     6241 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     6301 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     6361 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     6421 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     6481 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     6541 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     6601 gttttggaac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     6661 gcaaatgggc
//
