LOCUS       sequence-178030-        7075 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 7075)
  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 7075)
  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 HSPA1L
FEATURES             Location/Qualifiers
     source          1..7075
                     /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(3027..3044)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3033..3257
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3390..3409)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3541..3588
                     /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             3596..4618
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3596..4618
                     /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    4748..4869
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4785..4804)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4839..4858
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4918..4934)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4918..4934)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4931..4953)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4942..4958
                     /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(4966..4972,4973..4990,4991..4996))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5011..5032
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5149..5166)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5320..5908)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5400..5419)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6079..6870,6871..6939))
                     /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     6702..6721
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6940..7044)
                     /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 ctagtccagt gtggtggaat tctgcagata tcaccatggc tactgccaag ggaatcgcca
     1081 taggcatcga cctgggcacc acctactcct gtgtgggggt gttccagcac ggcaaggtgg
     1141 agatcatcgc caacgaccag ggcaaccgca ccacccccag ctacgtggcc ttcacagaca
     1201 ccgagcggct cattggggat gcggccaaga accaggtagc aatgaatccc cagaacactg
     1261 tttttgatgc taaacgtctg atcggcagga aatttaatga tcctgttgta caagcagata
     1321 tgaaactttg gccttttcaa gtgattaatg aaggaggcaa gcccaaagtc cttgtgtcct
     1381 acaaagggga gaataaagct ttctaccctg aggaaatctc ttcgatggta ttgactaagt
     1441 tgaaggagac tgctgaggcc tttttgggcc accctgtcac caatgcagtg attaccgtgc
     1501 cagcctattt caatgactct caacgtcagg ctactaagga tgcaggtgtg attgctggac
     1561 ttaatgtgct aagaatcatc aatgagccca cggctgctgc cattgcctat ggtttagata
     1621 aaggaggtca aggagaacga catgtcctga tttttgatct gggtggaggc acatttgatg
     1681 tgtcaattct gaccatagat gatgggattt ttgaggtaaa ggccactgct ggggacactc
     1741 acctgggtgg ggaggacttt gacaacaggc ttgtgagcca cttcgtggag gagttcaaga
     1801 ggaaacacaa aaaggacatc agccagaaca agcgagccgt gaggcggctg cgcaccgcct
     1861 gcgagagggc caagaggacc ctgtcgtcca gcacccaggc caacctagaa attgattcac
     1921 tttatgaagg cattgacttc tatacatcca tcaccagagc tcgatttgaa gagttgtgtg
     1981 cagacctgtt taggggtacc ctggagcctg tagaaaaagc gcttcgggat gccaagatgg
     2041 ataaggctaa aatccatgac attgttttag tagggggctc cacccgcatc cccaaggtgc
     2101 agcggctgct tcaggactac ttcaatggac gtgatctcaa caagagcatc aaccctgatg
     2161 aggccgtagc atatggggct gcggtacaag cagccatcct gatgggggac aagtctgaga
     2221 aggtacagga cctgctgctg ctggacgtgg ctcccctgtc cctggggctg gagacggctg
     2281 ggggcgtgat gactgccctg ataaagcgca actccaccat ccccaccaag cagacacaga
     2341 ttttcaccac ctactctgac aaccaacccg gggtgctgat ccaggtgtat gagggcgaga
     2401 gggccatgac aaaggacaac aacctgctgg ggcggtttga cctgactgga atccctccag
     2461 cacccagggg agttcctcag atcgaggtga cgtttgacat tgatgccaat ggtattctca
     2521 atgtcacagc cacggacaag agcaccggca aggtgaacaa gatcaccatc accaatgaca
     2581 agggccgcct gagcaaggag gagattgagc gcatggttct ggatgctgag aaatataaag
     2641 ctgaagatga ggtccagagg gagaaaattg ctgcaaagaa tgccttagaa tcctatgctt
     2701 ttaacatgaa gagtgttgtg agtgatgaag gtttgaaggg caagattagt gagtctgata
     2761 aaaataaaat attggataaa tgcaacgagc tcctttcgtg gctggaggtc aatcaactgg
     2821 cagagaaaga tgagtttgat cataagagaa aggaattgga gcagatgtgt aaccctatca
     2881 tcacaaaact ctaccaagga ggatgcactg ggcctgcctg cggaacaggg tatgtgcctg
     2941 gaaggcctgc cacaggcccc cacaattgaa gaagtagatt aagcggccgc tcgagtctag
     3001 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     3061 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     3121 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     3181 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     3241 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     3301 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3361 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3421 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     3481 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtaccta
     3541 gaagttccta ttccgaagtt cctattctct agaaagtata ggaacttcct tggccaaaaa
     3601 gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc
     3661 cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg
     3721 gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt
     3781 ttatcggcac tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggaatt
     3841 cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct
     3901 gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc
     3961 tgcggccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca
     4021 atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca
     4081 aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct
     4141 ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa
     4201 tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg
     4261 ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga
     4321 gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg
     4381 ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt
     4441 cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac
     4501 tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga
     4561 agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaatagca
     4621 cgtactacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4681 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4741 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4801 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4861 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4921 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4981 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5041 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5101 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5161 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5221 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5281 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5341 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5401 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5461 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5521 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5581 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5641 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5701 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5761 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5821 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5881 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5941 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6001 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6061 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6121 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6181 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6241 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6301 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6361 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6421 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6481 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6541 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6601 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6661 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6721 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6781 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6841 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6901 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6961 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7021 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
