LOCUS       Exported                6642 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA5/FRT/TO GFP DNAJB2a
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6642)
  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 6642)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6642
                     /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(1616..1633)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1622..1846
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1979..1998)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    2130..2177
                     /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             2185..3207
                     /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    3337..3458
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3374..3393)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3428..3447
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3507..3523)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3507..3523)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3520..3542)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3531..3547
                     /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(3555..3585)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3600..3621
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3738..3755)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3909..4497)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3989..4008)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4668..5528)
                     /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     5291..5310
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5529..5633)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5708..5727)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5899..6278
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6279..6482
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6433..6453
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    6484..6502
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6505..6523
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     6533..6557
                     /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 atgtgttctg agcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg
     1621 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     1681 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     1741 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     1801 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     1861 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     1921 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     1981 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     2041 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     2101 cttgattagg gtgatggttc acgtacctag aagttcctat tccgaagttc ctattctcta
     2161 gaaagtatag gaacttcctt ggccaaaaag cctgaactca ccgcgacgtc tgtcgagaag
     2221 tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa
     2281 tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc
     2341 gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg
     2401 attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc
     2461 cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag
     2521 ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc
     2581 ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg
     2641 attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc
     2701 gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac
     2761 ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg
     2821 gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc
     2881 ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat
     2941 ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa
     3001 ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc
     3061 gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc
     3121 gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc
     3181 agcactcgtc cgagggcaaa ggaatagcac gtactacgag atttcgattc caccgccgcc
     3241 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     3301 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     3361 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3421 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     3481 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     3541 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     3601 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     3661 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     3721 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3781 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3841 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3901 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3961 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4021 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4081 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4141 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4201 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4261 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4321 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4381 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4441 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4501 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4561 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4621 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4681 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4741 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4801 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4861 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4921 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4981 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5041 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5101 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5161 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5221 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5281 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5341 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5401 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5461 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5521 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5581 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5641 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
     5701 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     5761 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     5821 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
     5881 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
     5941 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     6001 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     6061 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     6121 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     6181 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     6241 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     6301 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     6361 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     6421 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     6481 ctctccctat cagtgataga gatctcccta tcagtgatag agatcgtcga cgagctcgtt
     6541 tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac
     6601 accgggaccg atccagcctc cggactctag cgtttaaact ta
//
