LOCUS       sequence-162060-        7767 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 7767)
  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 7767)
  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 GFP HSPA1L
FEATURES             Location/Qualifiers
     source          1..7767
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..3,4..6,7..717)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1..3,4..6,7..717)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(46..67)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(307..326)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     654..675
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2730..2747)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2736..2960
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3093..3112)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3244..3291
                     /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             3299..4321
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3299..4321
                     /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    4451..4572
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4488..4507)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4542..4561
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4621..4637)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4621..4637)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4634..4656)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4645..4661
                     /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(4669..4675,4676..4693,4694..4699))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4714..4735
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4852..4869)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5023..5611)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5103..5122)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5782..6573,6574..6642))
                     /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     6405..6424
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6643..6747)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6822..6841)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        7013..7392
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7393..7596
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7547..7567
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    7598..7616
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7619..7637
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     7647..7671
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      join(7762..7767,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
       61 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      121 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      181 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
      241 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      301 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      361 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      421 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
      481 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
      541 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      601 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
      661 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaaggga
      721 tccactagtc cagtgtggtg gaattctgca gatatcacca tggctactgc caagggaatc
      781 gccataggca tcgacctggg caccacctac tcctgtgtgg gggtgttcca gcacggcaag
      841 gtggagatca tcgccaacga ccagggcaac cgcaccaccc ccagctacgt ggccttcaca
      901 gacaccgagc ggctcattgg ggatgcggcc aagaaccagg tagcaatgaa tccccagaac
      961 actgtttttg atgctaaacg tctgatcggc aggaaattta atgatcctgt tgtacaagca
     1021 gatatgaaac tttggccttt tcaagtgatt aatgaaggag gcaagcccaa agtccttgtg
     1081 tcctacaaag gggagaataa agctttctac cctgaggaaa tctcttcgat ggtattgact
     1141 aagttgaagg agactgctga ggcctttttg ggccaccctg tcaccaatgc agtgattacc
     1201 gtgccagcct atttcaatga ctctcaacgt caggctacta aggatgcagg tgtgattgct
     1261 ggacttaatg tgctaagaat catcaatgag cccacggctg ctgccattgc ctatggttta
     1321 gataaaggag gtcaaggaga acgacatgtc ctgatttttg atctgggtgg aggcacattt
     1381 gatgtgtcaa ttctgaccat agatgatggg atttttgagg taaaggccac tgctggggac
     1441 actcacctgg gtggggagga ctttgacaac aggcttgtga gccacttcgt ggaggagttc
     1501 aagaggaaac acaaaaagga catcagccag aacaagcgag ccgtgaggcg gctgcgcacc
     1561 gcctgcgaga gggccaagag gaccctgtcg tccagcaccc aggccaacct agaaattgat
     1621 tcactttatg aaggcattga cttctataca tccatcacca gagctcgatt tgaagagttg
     1681 tgtgcagacc tgtttagggg taccctggag cctgtagaaa aagcgcttcg ggatgccaag
     1741 atggataagg ctaaaatcca tgacattgtt ttagtagggg gctccacccg catccccaag
     1801 gtgcagcggc tgcttcagga ctacttcaat ggacgtgatc tcaacaagag catcaaccct
     1861 gatgaggccg tagcatatgg ggctgcggta caagcagcca tcctgatggg ggacaagtct
     1921 gagaaggtac aggacctgct gctgctggac gtggctcccc tgtccctggg gctggagacg
     1981 gctgggggcg tgatgactgc cctgataaag cgcaactcca ccatccccac caagcagaca
     2041 cagattttca ccacctactc tgacaaccaa cccggggtgc tgatccaggt gtatgagggc
     2101 gagagggcca tgacaaagga caacaacctg ctggggcggt ttgacctgac tggaatccct
     2161 ccagcaccca ggggagttcc tcagatcgag gtgacgtttg acattgatgc caatggtatt
     2221 ctcaatgtca cagccacgga caagagcacc ggcaaggtga acaagatcac catcaccaat
     2281 gacaagggcc gcctgagcaa ggaggagatt gagcgcatgg ttctggatgc tgagaaatat
     2341 aaagctgaag atgaggtcca gagggagaaa attgctgcaa agaatgcctt agaatcctat
     2401 gcttttaaca tgaagagtgt tgtgagtgat gaaggtttga agggcaagat tagtgagtct
     2461 gataaaaata aaatattgga taaatgcaac gagctccttt cgtggctgga ggtcaatcaa
     2521 ctggcagaga aagatgagtt tgatcataag agaaaggaat tggagcagat gtgtaaccct
     2581 atcatcacaa aactctacca aggaggatgc actgggcctg cctgcggaac agggtatgtg
     2641 cctggaaggc ctgccacagg ccccacaatt gaagaagtag attaagcggc cgctcgagtc
     2701 tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     2761 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     2821 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     2881 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     2941 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     3001 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3061 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3121 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3181 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtac
     3241 ctagaagttc ctattccgaa gttcctattc tctagaaagt ataggaactt ccttggccaa
     3301 aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt
     3361 ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg
     3421 agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta
     3481 tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga
     3541 attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga
     3601 cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca tggatgcgat
     3661 cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg
     3721 tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg
     3781 gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat
     3841 gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa
     3901 caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt
     3961 cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat
     4021 ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct
     4081 ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa
     4141 tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg
     4201 gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt
     4261 agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata
     4321 gcacgtacta cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4381 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4441 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4501 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4561 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     4621 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4681 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     4741 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4801 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4861 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4921 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4981 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5041 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5101 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5161 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5221 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5281 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5341 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5401 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5461 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5521 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5581 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5641 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5701 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5761 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5821 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5881 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5941 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6001 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6061 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6121 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6181 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6241 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6301 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6361 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6421 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6481 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6541 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6601 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6661 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6721 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     6781 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
     6841 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     6901 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     6961 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
     7021 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     7081 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     7141 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     7201 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     7261 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     7321 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     7381 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     7441 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     7501 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     7561 aggcgtgtac ggtgggaggt ctatataagc agagctctcc ctatcagtga tagagatctc
     7621 cctatcagtg atagagatcg tcgacgagct cgtttagtga accgtcagat cgcctggaga
     7681 cgccatccac gctgttttga cctccataga agacaccggg accgatccag cctccggact
     7741 ctagcgttta aacttaagct tgccacc
//
