LOCUS       sequence-166280-        7636 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 7636)
  AUTHORS   Miao RQ, Fontana J, Fulton D, Lin MI, Harrison KD, Sessa WC
  TITLE     Dominant-negative Hsp90 reduces VEGF-stimulated nitric oxide release
            and migration in endothelial cells.
  JOURNAL   Arterioscler Thromb Vasc Biol. 2008 Jan . 28(1):105-11.
  PUBMED    17975117
REFERENCE   2  (bases 1 to 7636)
  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: 
            "Arterioscler Thromb Vasc Biol. 2008 Jan . 28(1):105-11."
COMMENT     Sequence Label: Dominant negative Hsp90 beta
FEATURES             Location/Qualifiers
     source          1..7636
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        140..519
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        520..723
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     674..694
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     768..787
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        768..786
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             883..909
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(3094..3112)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3095..3112)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3132..3149)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3138..3362
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3408..3836
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3495..3514)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3705..3726
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3845..3865)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3850..4179
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4030..4165
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4092..4111
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4246..5040
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4300..4319)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4910..4929
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5214..5335
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5251..5270)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5305..5324
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5384..5400)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5384..5400)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5397..5419)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5408..5424
                     /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(5432..5438,5439..5456,5457..5462))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5477..5498
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5615..5632)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5786..6374)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5866..5885)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6545..7336,7337..7405))
                     /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     7168..7187
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7406..7510)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7585..7604)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
       61 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
      121 cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg
      181 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      241 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      301 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      361 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      421 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      481 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
      541 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
      601 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
      661 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
      721 gctctctggc taactagaga acccactgct tactggctta tcgaaattaa tacgactcac
      781 tatagggaga cccaagcttg gtaccgagct cggatccact agtaacggcc gccagtgtgc
      841 tggaattctg cagatatcca tcacactggc ggccgcgcca tgtacccata cgacgtccca
      901 gactacgctg aggaagtgca ccatggagag gaggaggtgg agacttttgc ctttcaggca
      961 gaaattgccc aactcatgtc cctcatcatc aataccttct attccaacaa ggagattttc
     1021 cttcgggagt tgatctctaa tgcttctgat gccttggaca agattcgcta tgagagcctg
     1081 acagaccctt cgaagttgga cagtggtaaa gagctgaaaa ttgacatcat ccccaaccct
     1141 caggaacgta ccctgacttt ggttaacaca ggcattggca tgaccaaagc tgatctcata
     1201 aataatttgg gaaccattgc caagtctggt actaaagcat tcatggaggc tcttcaggct
     1261 ggtgcagaca tctccatgat tgggcagttt ggtgttggct tttattctgc ctacttggtg
     1321 gcagagaaag tggttgtgat cacaaagcac aacgatgatg aacagtatgc ttgggagtct
     1381 tctgctggag gttccttcac tgtgcgtgct gaccatggtg agcccattgg caggggtacc
     1441 aaagtgatcc tccatcttaa agaagatcag acagagtacc tagaagagag gcgggtcaaa
     1501 gaagtagtga agaagcattc tcagttcata ggctatccca tcacccttta tttggagaag
     1561 gaacgagaga aggaaattag tgatgatgag gcagaggaag agaaaggtga gaaagaagag
     1621 gaagataaag atgatgaaga aaagcccaag atcgaagatg tgggttcaga tgaggaggat
     1681 gacagcggta aggataagaa gaagaaaact aagaagatca aagagaaata cattgatcag
     1741 gaagaactaa acaagaccaa gcctatttgg accagaaacc ctgatgacat cacccaagag
     1801 gagtatggag aattctacaa gagcctcact aatgactggg aagaccactt ggcagtcaag
     1861 cacttttctg tagaaggtca gttggaattc agggcattgc tatttattcc tcgtcgggct
     1921 ccctttgacc tttttgagaa caagaagaaa aagaacaaca tcaaactcta tgtccgccgt
     1981 gtgttcatca tggacagctg tgatgagttg ataccagagt atctcaattt tatccgtggt
     2041 gtggttgact ctgaggatct gcccctgaac atctcccgag aaatgctcca gcagagcaaa
     2101 atcttgaaag tcattcgcaa aaacattgtt aagaagtgcc ttgagctctt ctctgagctg
     2161 gcagaagaca aggagaatta caagaaattc tatgaggcat tctctaaaaa tctcaagctt
     2221 ggaatccacg aagactccac taaccgccgc cgcctgtctg agctgctgcg ctatcatacc
     2281 tcccagtctg gagatgagat gacatctctg tcagagtatg tttctcgcat gaaggagaca
     2341 cagaagtcca tctattacat cactggtgag agcaaagagc aggtggccaa ctcagctttt
     2401 gtggagcgag tgcggaaacg gggcttcgag gtggtatata tgaccgagcc cattgacgag
     2461 tactgtgtgc agcagctcaa ggaatttgat gggaagagcc tggtctcagt taccaaggag
     2521 ggtctggagc tgcctgagga tgaggaggag aagaagaaga tggaagagag caaggcaaag
     2581 tttgagaacc tctgcaagct catgaaagaa atcttagata agaaggttga gaaggtgaca
     2641 atctccaata gacttgtgtc ttcaccttgc tgcattgtga ccagcaccta cggctggaca
     2701 gccaatatgg agcggatcat gaaagcccag gcacttcggg acaactccac catgggctat
     2761 atgatggcca aaaagcacct ggagatcaac cctgaccacc ccattgtgga gacgctgcgg
     2821 cagaaggctg aggccgacaa gaatgataag gcagttaagg acctggtggt gctgctgttt
     2881 gaaaccgccc tgctatcttc tggcttttcc cttgaggatc cccagaccca ctccaaccgc
     2941 atctatcgca tgatcaagct aggtctaggt attgatgaag atgaagtggc agcagaggaa
     3001 cccaatgctg cagttcctga tgagatcccc cctctcgagg gcgatgagga tgcgtctcgc
     3061 atggaagaag tcgattagtc tagagggccc tattctatag tgtcacctaa atgctagagc
     3121 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3181 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3241 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3301 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3361 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     3421 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3481 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3541 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3601 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3661 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3721 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     3781 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3841 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3901 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3961 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4021 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4081 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4141 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4201 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4261 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4321 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4381 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4441 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4501 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4561 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4621 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4681 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4741 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4801 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4861 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4921 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4981 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5041 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5101 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5161 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     5221 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5281 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5341 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     5401 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5461 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5521 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5581 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5641 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5701 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5761 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5821 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5881 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5941 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6001 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6061 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6121 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6181 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6241 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6301 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6361 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6421 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6481 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6541 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6601 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6661 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6721 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6781 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6841 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6901 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6961 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7021 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7081 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7141 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7201 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7261 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7321 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7381 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7441 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7501 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     7561 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     7621 tctgctccct gcttgt
//
