LOCUS       sequence-202011-        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   Garcia-Cardena G, Fan R, Shah V, Sorrentino R, Cirino G, 
            Papapetropoulos A, Sessa WC
  TITLE     Dynamic activation of endothelial nitric oxide synthase by Hsp90.
  JOURNAL   Nature. 1998 Apr 23. 392(6678):821-4.
  PUBMED    9580552
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: "Nature. 
            1998 Apr 23. 392(6678):821-4."
COMMENT     Sequence Label: Hsp90 HA
FEATURES             Location/Qualifiers
     source          1..7636
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     8..25
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    142..163
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(178..183,184..201,202..208)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    216..232
                     /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)."
     primer_bind     221..243
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     240..256
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     240..256
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    305..426
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(316..335)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     370..389
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(600..1394)
                     /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(711..730)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1321..1340
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1461..1790)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1475..1610
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1529..1548)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     1775..1795
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(1804..2232)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1914..1935)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2126..2145
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2278..2502
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2491..2508
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        2528..2546
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2528..2545
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(4731..4757)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4853..4872)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4854..4872)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4917..5120)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4946..4966)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5121..5500
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     5672..5691
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5766..5870
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5871..5939,5940..6731)
                     /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(6089..6108)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6902..7490
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7391..7410
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
       61 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
      121 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
      181 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
      241 aggaaacagc tatgaccatg attacgccaa gctctagcta gaggtcgacg gtatacagac
      301 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
      361 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
      421 caagttgggg tgggcgaaga actccagcat gagatccccg cgctggagga tcatccagcc
      481 ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa
      541 atctcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct
      601 cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata
      661 ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg
      721 gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat
      781 ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg
      841 acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg cgaacagttc ggctggcgcg
      901 agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta
      961 cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc
     1021 gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga
     1081 gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca
     1141 gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc
     1201 gctgcctcgt cctgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc
     1261 gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt
     1321 gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca
     1381 tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatcagatcc gaaaatggat
     1441 atacaagctc ccgggagctt tttgcaaaag cctaggcctc caaaaaagcc tcctcactac
     1501 ttctggaata gctcagaggc agaggcggcc tcggcctctg cataaataaa aaaaattagt
     1561 cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt
     1621 aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct
     1681 ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata
     1741 cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac attccacaga
     1801 attaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     1861 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     1921 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     1981 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     2041 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     2101 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     2161 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     2221 tacagggcgc gtggggatac cccctagagc cccagctggt tctttccgcc tcagaagcca
     2281 tagagcccac cgcatcccca gcatgcctgc tattgtcttc ccaatcctcc cccttgctgt
     2341 cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc gatgcaattt
     2401 cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag gaaggcacgg
     2461 gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaggct gatcagcgag
     2521 ctctagcatt taggtgacac tatagaatag ggccctctag actaatcgac ttcttccatg
     2581 cgagacgcat cctcatcgcc ctcgagaggg gggatctcat caggaactgc agcattgggt
     2641 tcctctgctg ccacttcatc ttcatcaata cctagaccta gcttgatcat gcgatagatg
     2701 cggttggagt gggtctgggg atcctcaagg gaaaagccag aagatagcag ggcggtttca
     2761 aacagcagca ccaccaggtc cttaactgcc ttatcattct tgtcggcctc agccttctgc
     2821 cgcagcgtct ccacaatggg gtggtcaggg ttgatctcca ggtgcttttt ggccatcata
     2881 tagcccatgg tggagttgtc ccgaagtgcc tgggctttca tgatccgctc catattggct
     2941 gtccagccgt aggtgctggt cacaatgcag caaggtgaag acacaagtct attggagatt
     3001 gtcaccttct caaccttctt atctaagatt tctttcatga gcttgcagag gttctcaaac
     3061 tttgccttgc tctcttccat cttcttcttc tcctcctcat cctcaggcag ctccagaccc
     3121 tccttggtaa ctgagaccag gctcttccca tcaaattcct tgagctgctg cacacagtac
     3181 tcgtcaatgg gctcggtcat atataccacc tcgaagcccc gtttccgcac tcgctccaca
     3241 aaagctgagt tggccacctg ctctttgctc tcaccagtga tgtaatagat ggacttctgt
     3301 gtctccttca tgcgagaaac atactctgac agagatgtca tctcatctcc agactgggag
     3361 gtatgatagc gcagcagctc agacaggcgg cggcggttag tggagtcttc gtggattcca
     3421 agcttgagat ttttagagaa tgcctcatag aatttcttgt aattctcctt gtcttctgcc
     3481 agctcagaga agagctcaag gcacttctta acaatgtttt tgcgaatgac tttcaagatt
     3541 ttgctctgct ggagcatttc tcgggagatg ttcaggggca gatcctcaga gtcaaccaca
     3601 ccacggataa aattgagata ctctggtatc aactcatcac agctgtccat gatgaacaca
     3661 cggcggacat agagtttgat gttgttcttt ttcttcttgt tctcaaaaag gtcaaaggga
     3721 gcccgacgag gaataaatag caatgccctg aattccaact gaccttctac agaaaagtgc
     3781 ttgactgcca agtggtcttc ccagtcatta gtgaggctct tgtagaattc tccatactcc
     3841 tcttgggtga tgtcatcagg gtttctggtc caaataggct tggtcttgtt tagttcttcc
     3901 tgatcaatgt atttctcttt gatcttctta gttttcttct tcttatcctt accgctgtca
     3961 tcctcctcat ctgaacccac atcttcgatc ttgggctttt cttcatcatc tttatcttcc
     4021 tcttctttct cacctttctc ttcctctgcc tcatcatcac taatttcctt ctctcgttcc
     4081 ttctccaaat aaagggtgat gggatagcct atgaactgag aatgcttctt cactacttct
     4141 ttgacccgcc tctcttctag gtactctgtc tgatcttctt taagatggag gatcactttg
     4201 gtacccctgc caatgggctc accatggtca gcacgcacag tgaaggaacc tccagcagaa
     4261 gactcccaag catactgttc atcatcgttg tgctttgtga tcacaaccac tttctctgcc
     4321 accaagtagg cagaataaaa gccaacacca aactgcccaa tcatggagat gtctgcacca
     4381 gcctgaagag cctccatgaa tgctttagta ccagacttgg caatggttcc caaattattt
     4441 atgagatcag ctttggtcat gccaatgcct gtgtcaacca aagtcagggt acgttcctga
     4501 gggttgggga tgatgtcaat tttcagctct ttaccactgt ccaacttcga agggtctgtc
     4561 aggctctcat agcgaatctt gtccaaggca tcagaagcat tagagatcaa ctcccgaagg
     4621 aaaatctcct tgttggaata gaaggtattg atgatgaggg acatgagttg ggcaatttct
     4681 gcctgaaagg caaaagtctc cacctcctcc tctccatggt gcacttcctc agcgtagtct
     4741 gggacgtcgt atgggtacat ggcgcggccg ccagtgtgat ggatatctgc agaattccag
     4801 cacactggcg gccgttacta gtggatccga gctcggtacc aagcttgggt ctccctatag
     4861 tgagtcgtat taatttcgat aagccagtaa gcagtgggtt ctctagttag ccagagagct
     4921 ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag
     4981 ttgttacgac attttggaaa gtcccgttga ttttggtgcc aaaacaaact cccattgacg
     5041 tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg cccattgatg
     5101 tactgccaaa accgcatcac catggtaata gcgatgacta atacgtagat gtactgccaa
     5161 gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca tttaccgtca
     5221 ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc caagtgggca
     5281 gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg cgttactatg
     5341 ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg
     5401 ccatttaccg taagttatgt aacgcggaac tccatatatg ggctatgaac taatgacccc
     5461 gtaattgatt actattaata actagtcaat aatcaatgtc aacgcgtata tctggcccgt
     5521 acatcgcgaa gcagcgcaaa acgcctaacc ctaagcagat tcttcatgca attgtcggtc
     5581 aagccttgcc ttgttgtagc ttaaattttg ctcgcgcact actcagcgac ctccaacaca
     5641 caagcaggga gcagatactg gcttaactat gcggcatcag agcagattgt actgagagtg
     5701 caccataggg gatcgggaga tctcccgatc cgtcgacgtc aggtggcact tttcggggaa
     5761 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5821 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     5881 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     5941 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     6001 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     6061 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     6121 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     6181 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     6241 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     6301 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     6361 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     6421 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     6481 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     6541 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     6601 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     6661 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     6721 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     6781 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6841 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6901 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6961 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     7021 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     7081 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     7141 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     7201 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     7261 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     7321 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     7381 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     7441 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     7501 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     7561 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     7621 gccgcagccg aacgac
//
