LOCUS       Exported                6783 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA5/FRT/TO GFP DNAJB2b
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6783)
  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 6783)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6783
                     /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"
     RBS             875..883
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(1757..1774)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1763..1987
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2120..2139)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    2271..2318
                     /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             2326..3348
                     /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    3478..3599
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3515..3534)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3569..3588
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3648..3664)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3648..3664)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3661..3683)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3672..3688
                     /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(3696..3726)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3741..3762
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3879..3896)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4050..4638)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4130..4149)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4809..5669)
                     /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     5432..5451
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5670..5774)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5849..5868)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6040..6419
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6420..6623
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6574..6594
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    6625..6643
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6646..6664
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     6674..6698
                     /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 atccaccatg gcatcctact acgagatcct agacgtgccg cgaagtgcgt
      781 ccgctgatga catcaagaag gcgtatcggc gcaaggctct ccagtggcac ccagacaaaa
      841 acccagataa taaagagttt gctgagaaga aatttaagga ggtggccgag gcatatgaag
      901 tgctgtctga caagcacaag cgggagattt acgaccgcta tggccgggaa gggctgacag
      961 ggacaggaac tggcccatct cgggcagaag ctggcagtgg tgggcctggc ttcaccttca
     1021 ccttccgcag ccccgaggag gtcttccggg aattctttgg gagtggagac ccttttgcag
     1081 agctctttga tgacctgggc cccttctcag agcttcagaa ccggggttcc cgacactcag
     1141 gccccttctt taccttctct tcctccttcc ctgggcactc cgatttctcc tcctcatctt
     1201 tctccttcag tcctggggct ggtgcttttc gctctgtttc tacatctacc acctttgtcc
     1261 aaggacgccg catcaccaca cgcagaatca tggagaacgg gcaggagcgg gtggaagtgg
     1321 aggaggatgg gcagctgaag tcagtcacaa tcaatggtgt cccagatgac ctggcactgg
     1381 gcttggagct gagccgtcgc gagcagcagc cgtcagtcac ttccaggtct gggggcactc
     1441 aggtccagca gacccctgcc tcatgcccct tggacagcga cctctctgag gatgaggacc
     1501 tgcagctggc catggcctac agcctgtcag agatggaggc agctgggaag aaacccgcag
     1561 gtgggcggga ggcacagcac cgacggcagg ggcggcccaa ggcccagcac caagatccag
     1621 gcttgggggg gacccaggag ggtgcgaggg gtgaagcaac caaacgcagt ccatccccag
     1681 aggagaaggc ctctcgctgc ctcatcctct gagcggccgc tcgagtctag agggcccgtt
     1741 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     1801 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     1861 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     1921 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     1981 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     2041 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2101 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2161 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2221 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtaccta gaagttccta
     2281 ttccgaagtt cctattctct agaaagtata ggaacttcct tggccaaaaa gcctgaactc
     2341 accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc cgacctgatg
     2401 cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg gcgtggatat
     2461 gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt ttatcggcac
     2521 tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggaatt cagcgagagc
     2581 ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc
     2641 gaactgcccg ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat
     2701 cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca atacactaca
     2761 tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca aactgtgatg
     2821 gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct ttgggccgag
     2881 gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg
     2941 gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg ggattcccaa
     3001 tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga gcagcagacg
     3061 cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg ggcgtatatg
     3121 ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt cgatgatgca
     3181 gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt
     3241 acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc
     3301 gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaatagca cgtactacga
     3361 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     3421 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     3481 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3541 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     3601 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     3661 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     3721 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     3781 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     3841 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3901 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3961 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4021 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4081 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4141 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4201 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4261 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4321 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4381 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4441 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4501 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4561 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4621 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4681 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4741 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4801 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4861 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4921 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4981 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5041 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5101 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5161 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5221 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5281 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5341 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5401 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5461 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5521 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5581 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5641 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5701 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5761 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     5821 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     5881 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
     5941 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
     6001 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
     6061 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     6121 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     6181 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     6241 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     6301 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     6361 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     6421 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     6481 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     6541 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     6601 gggaggtcta tataagcaga gctctcccta tcagtgatag agatctccct atcagtgata
     6661 gagatcgtcg acgagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct
     6721 gttttgacct ccatagaaga caccgggacc gatccagcct ccggactcta gcgtttaaac
     6781 tta
//
