LOCUS       sequence-9152-ex        7068 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 7068)
  AUTHORS   Hageman J, Kampinga HH
  TITLE     Computational analysis of the human HSPH/HSPA/DNAJ family and 
            cloning of a human HSPH/HSPA/DNAJ expression library.
  JOURNAL   Cell Stress Chaperones. 2009 Jan;14(1):1-21. doi: 
            10.1007/s12192-008-0060-2. Epub 2008 Aug 7.
  PUBMED    18686016
REFERENCE   2  (bases 1 to 7068)
  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: 
            "10.1007/s12192-008-0060-2"; journalName: "Cell Stress Chaperones"; 
            date: "2009-01"; volume: "14"; issue: "1"; pages: "1-21"
COMMENT     Sequence Label: pcDNA5/FRT/TO V5 SSA1
FEATURES             Location/Qualifiers
     source          1..7068
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             996..1037
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3020..3037)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3026..3250
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3383..3402)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3534..3581
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3589..4611
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3589..4611
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4741..4862
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4778..4797)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4832..4851
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4911..4927)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4911..4927)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4924..4946)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4935..4951
                     /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(4959..4965,4966..4983,4984..4989))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5004..5025
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5142..5159)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5313..5901)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5393..5412)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6072..6863,6864..6932))
                     /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     6695..6714
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6933..7037)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttgaattca ccatgggtaa gcctatccct aaccctctcc
     1021 tcggtctcga ttctacggga tccaccatgt caaaagctgt cggtattgat ttaggtacaa
     1081 catactcgtg tgttgctcac tttgctaatg atcgtgtgga cattattgcc aacgatcaag
     1141 gtaacagaac cactccatct tttgtcgctt tcactgacac tgaaagattg attggtgatg
     1201 ctgctaagaa tcaagctgct atgaatcctt cgaataccgt tttcgacgct aagcgtttga
     1261 tcggtagaaa cttcaacgac ccagaagtgc aggctgacat gaagcacttc ccattcaagt
     1321 tgatcgatgt tgacggtaag cctcaaattc aagttgaatt taagggtgaa accaagaact
     1381 ttaccccaga acaaatctcc tccatggtct tgggtaagat gaaggaaact gccgaatctt
     1441 acttgggagc caaggtcaat gacgctgtcg tcactgtccc agcttacttc aacgattctc
     1501 aaagacaagc taccaaggat gctggtacca ttgctggttt gaatgtcttg cgtattatta
     1561 acgaacctac cgccgctgcc attgcttacg gtttggacaa gaagggtaag gaagaacacg
     1621 tcttgatttt cgacttgggt ggtggtactt tcgatgtctc tttgttgttc attgaagacg
     1681 gtatctttga agttaaggcc accgctggtg acacccattt gggtggtgaa gattttgaca
     1741 acagattggt caaccacttc atccaagaat tcaagagaaa gaacaagaag gacttgtcta
     1801 ccaaccaaag agctttgaga agattaagaa ccgcttgtga aagagccaag agaactttgt
     1861 cttcctccgc tcaaacttcc gttgaaattg actctttgtt cgaaggtatc gatttctaca
     1921 cttccatcac cagagccaga ttcgaagaat tgtgtgctga cttgttcaga tctactttgg
     1981 acccagttga aaaggtcttg agagatgcta aattggacaa atctcaagtc gatgaaattg
     2041 tcttggtcgg tggttctacc agaattccaa aggtccaaaa attggtcact gactacttca
     2101 acggtaagga accaaacaga tctatcaacc cagatgaagc tgttgcttac ggtgctgctg
     2161 ttcaagctgc tattttgact ggtgacgaat cttccaagac tcaagatcta ttgttgttgg
     2221 atgtcgctcc attatccttg ggtattgaaa ctgctggtgg tgtcatgacc aagttgattc
     2281 caagaaactc taccatttca acaaagaagt tcgagatctt ttccacttat gctgataacc
     2341 aaccaggtgt cttgattcaa gtctttgaag gtgaaagagc caagactaag gacaacaact
     2401 tgttgggtaa gttcgaattg agtggtattc caccagctcc aagaggtgtc ccacaaattg
     2461 aagtcacttt cgatgtcgac tctaacggta ttttgaatgt ttccgccgtc gaaaagggta
     2521 ctggtaagtc taacaagatc actattacca acgacaaggg tagattgtcc aaggaagata
     2581 tcgaaaagat ggttgctgaa gccgaaaaat tcaaggaaga agatgaaaag gaatctcaaa
     2641 gaattgcttc caagaaccaa ttggaatcca ttgcttactc tttgaagaac accatttctg
     2701 aagctggtga caaattggaa caagctgaca aggacaccgt caccaagaag gctgaagaga
     2761 ctatttcttg gttagacagc aacaccactg ccagcaagga agaattcgat gacaagttga
     2821 aggagttgca agacattgcc aacccaatca tgtctaagtt gtaccaagct ggtggtgctc
     2881 caggtggcgc tgcaggtggt gctccaggcg gtttcccagg tggtgctcct ccagctccag
     2941 aggctgaagg tccaaccgtt gaagaagttg attaagcggc cgctcgagtc tagagggccc
     3001 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     3061 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     3121 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     3181 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     3241 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     3301 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3361 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3421 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3481 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtac ctagaagttc
     3541 ctattccgaa gttcctattc tctagaaagt ataggaactt ccttggccaa aaagcctgaa
     3601 ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg
     3661 atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg agggcgtgga
     3721 tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg
     3781 cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga attcagcgag
     3841 agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa
     3901 accgaactgc ccgctgttct gcagccggtc gcggaggcca tggatgcgat cgctgcggcc
     3961 gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg tcaatacact
     4021 acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg gcaaactgtg
     4081 atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat gctttgggcc
     4141 gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa caatgtcctg
     4201 acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt cggggattcc
     4261 caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat ggagcagcag
     4321 acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct ccgggcgtat
     4381 atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa tttcgatgat
     4441 gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg gactgtcggg
     4501 cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt agaagtactc
     4561 gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata gcacgtacta
     4621 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4681 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     4741 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4801 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4861 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     4921 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4981 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5041 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5101 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5161 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5221 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5281 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5341 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5401 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5461 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5521 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5581 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5641 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5701 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5761 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5821 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5881 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5941 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6001 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6061 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6121 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6181 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6241 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6301 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6361 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6421 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6481 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6541 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6601 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6661 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6721 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6781 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6841 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6901 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6961 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7021 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
