LOCUS       sequence-178033-        7035 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 7035)
  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 7035)
  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 HIS HSPA2
FEATURES             Location/Qualifiers
     source          1..7035
                     /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..1013
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2987..3004)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2993..3217
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3350..3369)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3501..3548
                     /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             3556..4578
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3556..4578
                     /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    4708..4829
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4745..4764)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4799..4818
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4878..4894)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4878..4894)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4891..4913)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4902..4918
                     /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(4926..4932,4933..4950,4951..4956))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4971..4992
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5109..5126)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5280..5868)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5360..5379)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6039..6830,6831..6899))
                     /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     6662..6681
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6900..7004)
                     /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 cttctgcaga ccatgcatca tcaccatcac catggatcca
     1021 ccatgtctgc ccgtggcccg gctatcggca tcgacctggg caccacctat tcgtgcgtcg
     1081 gggtcttcca acatggcaag gtggagatca tcgccaacga ccagggcaat cgcaccaccc
     1141 ccagctacgt ggccttcacg gacaccgagc gcctcatcgg cgacgccgcc aagaaccagg
     1201 tggccatgaa ccccaccaac accatcttcg acgccaagag gctgattgga cggaaattcg
     1261 aggatgccac agtgcagtcg gatatgaaac actggccgtt ccgggtggtg agcgagggag
     1321 gcaagcccaa agtgcaagta gagtacaagg gggagaccaa gaccttcttc ccagaggaga
     1381 tatcctccat ggtcctcacg aagatgaagg agatcgcgga agcctacctg gggggcaagg
     1441 tgcacagcgc ggtcataacg gtcccggcct atttcaacga ctcgcagcgc caggccacca
     1501 aggacgcagg caccatcacg gggctcaatg tgctgcgcat catcaacgag cccacggcgg
     1561 cggccatcgc ctacggcctg gacaagaagg gctgcgcggg cggcgagaag aacgtgctca
     1621 tctttgacct gggcggtggc actttcgacg tgtccatcct gaccatcgag gatggcatct
     1681 tcggggtgaa gtccacggcc ggcgataccc acctgggcgg tgaggacttc gacaaccgca
     1741 tggtgagcca cctggcggag gagttcaagc gcaagcacaa gaaggacatt gggcccaaca
     1801 agcgcgccgt gaggcggctg cgcaccgctt gcgagcgcgc caagcgcacc ctgagctcgt
     1861 ccacgcaggc gagcatcgag atcgactcgc tctacgaggg cgtggacttc tatacgtcca
     1921 tcacgcgcgc ccgcttcgag gagctcaatg ccgacctctt tcgcgggacc ctggagccgg
     1981 tggagaaggc gctgcgcgac gccaagctgg acaagggcca gatccaggag atcgtgctgg
     2041 tgggcggctc cactcgtatc cccaagatcc agaagctgct gcaggatttc ttcaacggca
     2101 aggagctgaa caagagcatc aaccccgacg aggcggtggc ctatggcgcc gcggtgcagg
     2161 cggccatcct catcggcgac aaatcagaga atgtgcagga cctgctgcta ctcgacgtga
     2221 ccccgttgtc gctgggcatc gagacagctg gcggtgtcat gaccccactc atcaagagga
     2281 acaccacgat ccccaccaag cagacgcaga ccttcaccac ctactcggac aaccagagca
     2341 gcgtactggt gcaggtatac gagggcgaac gggccatgac caaggacaat aacctgctgg
     2401 gcaagttcga cctgaccggg attccccctg cgcctcgcgg ggtcccccaa atcgaggtta
     2461 ccttcgacat tgacgccaat ggcatcctta acgttaccgc cgccgacaag agcaccggta
     2521 aggaaaacaa aatcaccatc accaatgaca aaggtcgtct gagcaaggac gacattgacc
     2581 ggatggtgca ggaggcggag cggtacaaat cggaagatga ggcgaatcgc gaccgagtcg
     2641 cggccaaaaa cgccctggag tcctatacct acaacatcaa gcagacggtg gaagacgaga
     2701 aactgagggg caagattagc gagcaggaca aaaacaagat cctcgacaag tgtcaggagg
     2761 tgatcaactg gctcgaccga aaccagatgg cagagaaaga tgagtatgaa cacaagcaga
     2821 aagagctcga aagagtttgc aaccccatca tcagcaaact ttaccaaggt ggtcctggcg
     2881 gcggcagcgg cggcggcggt tcaggagcct ccgggggacc caccatcgaa gaagtggact
     2941 aagcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct
     3001 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3061 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3121 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3181 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     3241 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3301 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3361 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3421 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3481 ggtgatggtt cacgtaccta gaagttccta ttccgaagtt cctattctct agaaagtata
     3541 ggaacttcct tggccaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc
     3601 gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct
     3661 ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt
     3721 ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa
     3781 gtgcttgaca ttggggaatt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag
     3841 ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg
     3901 gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc
     3961 ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat
     4021 ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag
     4081 gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac
     4141 gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac
     4201 tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg
     4261 ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt
     4321 gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag
     4381 agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc
     4441 gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc
     4501 tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt
     4561 ccgagggcaa aggaatagca cgtactacga gatttcgatt ccaccgccgc cttctatgaa
     4621 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4681 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     4741 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4801 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     4861 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4921 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4981 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5041 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5101 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5161 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5221 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5281 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5341 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5401 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5461 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5521 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5581 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5641 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5701 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5761 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5821 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5881 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5941 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6001 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6061 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6121 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6181 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6241 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6301 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6361 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6421 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6481 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6541 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6601 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6661 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6721 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6781 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6841 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6901 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6961 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     7021 gtgccacctg acgtc
//
