LOCUS       Exported                6537 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA5/FRT/TO GFP DNAJB6b
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6537)
  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. 2008 Aug 7. ():.
  PUBMED    18686016
REFERENCE   2  (bases 1 to 6537)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6537
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     regulatory      6..15
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             12..728
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             12..728
                     /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(57..78)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(318..337)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     665..686
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1511..1528)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1517..1741
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1874..1893)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    2025..2072
                     /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             2080..3102
                     /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    3232..3353
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3269..3288)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3323..3342
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3402..3418)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3402..3418)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3415..3437)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3426..3442
                     /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(3450..3480)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3495..3516
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3633..3650)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3804..4392)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3884..3903)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4563..5423)
                     /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     5186..5205
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5424..5528)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5603..5622)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5794..6173
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6174..6377
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6328..6348
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    6379..6397
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6400..6418
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     6428..6452
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 agcttgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
       61 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
      121 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
      181 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
      241 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
      301 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
      361 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
      421 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
      481 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
      541 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
      601 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
      661 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
      721 tgtacaaggg atccggaacc atggtggatt actatgaagt tctaggcgtg cagagacatg
      781 cctcacccga ggatattaaa aaggcatatc ggaaactggc actgaagtgg catccagata
      841 aaaatcctga gaataaagaa gaagcagaga gaaaattcaa gcaagtagcg gaggcatatg
      901 aagtgctgtc ggatgctaag aaacgggaca tctatgacaa atatggcaaa gaaggattaa
      961 atggtggagg aggaggtgga agtcattttg acagtccatt tgaatttggc ttcacattcc
     1021 gtaacccaga tgatgtcttc agggaatttt ttggtggaag ggacccattt tcatttgact
     1081 tctttgaaga cccttttgag gacttctttg ggaatcgaag gggtccccga ggaagcagaa
     1141 gccgagggac ggggtcgttt ttctctgcgt tcagtggatt tccgtctttt ggaagtggat
     1201 tttcttcttt tgatacagga tttacttcat ttgggtcact aggtcacggg ggcctcactt
     1261 cattctcttc cacgtcattt ggtggtagtg gcatgggcaa cttcaaatcg atatcaactt
     1321 caactaaaat ggttaatggc agaaaaatca ctacaaagag aattgtcgag aacggtcaag
     1381 aaagagtaga agttgaagaa gatggccagt taaagtcctt aacaataaat ggtaaggagc
     1441 agctgctgcg cttggataac aagtaagcgg ccgctcgagt ctagagggcc cgtttaaacc
     1501 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     1561 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     1621 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     1681 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     1741 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     1801 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     1861 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     1921 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     1981 ctcgacccca aaaaacttga ttagggtgat ggttcacgta cctagaagtt cctattccga
     2041 agttcctatt ctctagaaag tataggaact tccttggcca aaaagcctga actcaccgcg
     2101 acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc
     2161 tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg
     2221 cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca
     2281 tcggccgcgc tcccgattcc ggaagtgctt gacattgggg aattcagcga gagcctgacc
     2341 tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg
     2401 cccgctgttc tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc
     2461 cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt
     2521 gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac
     2581 accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc
     2641 cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat
     2701 ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag
     2761 gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac
     2821 ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc
     2881 attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg
     2941 gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa
     3001 atcgcccgca gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt
     3061 ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat agcacgtact acgagatttc
     3121 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     3181 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     3241 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3301 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     3361 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     3421 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     3481 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     3541 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     3601 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3661 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3721 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3781 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3841 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3901 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3961 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4021 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4081 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4141 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4201 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4261 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4321 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4381 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4441 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4501 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4561 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4621 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4681 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4741 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4801 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4861 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4921 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4981 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5041 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5101 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5161 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5221 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5281 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5341 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5401 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5461 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5521 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg
     5581 atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     5641 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     5701 caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc
     5761 tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact agttattaat
     5821 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     5881 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     5941 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     6001 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     6061 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     6121 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     6181 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     6241 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     6301 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     6361 tctatataag cagagctctc cctatcagtg atagagatct ccctatcagt gatagagatc
     6421 gtcgacgagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg
     6481 acctccatag aagacaccgg gaccgatcca gcctccggac tctagcgttt aaactta
//
