LOCUS       Exported                7734 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA5/FRT/TO GFP HSPA1A
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7734)
  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 7734)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7734
                     /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(2708..2725)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2714..2938
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3071..3090)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3222..3269
                     /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             3277..4299
                     /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    4429..4550
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4466..4485)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4520..4539
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4599..4615)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4599..4615)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4612..4634)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4623..4639
                     /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(4647..4677)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4692..4713
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4830..4847)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5001..5589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5081..5100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5760..6620)
                     /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     6383..6402
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6621..6725)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6800..6819)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6991..7370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7371..7574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7525..7545
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    7576..7594
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7597..7615
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     7625..7649
                     /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 gccaaagccg cggcgatcgg catcgacctg ggcaccacct
      781 actcctgcgt gggggtgttc caacacggca aggtggagat catcgccaac gaccagggca
      841 accgcaccac ccccagctac gtggccttca cggacaccga gcggctcatc ggggatgcgg
      901 ccaagaacca ggtggcgctg aacccgcaga acaccgtgtt tgacgcgaag cggctgatcg
      961 gccgcaagtt cggcgacccg gtggtgcagt cggacatgaa gcactggcct ttccaggtga
     1021 tcaacgacgg agacaagccc aaggtgcagg tgagctacaa gggggacacc aaggcattct
     1081 accccgagga gatctcgtcc atggtgctga ccaagatgaa ggagatcgcc gaggcgtacc
     1141 tgggctaccc ggtgaccaac gcggtgatca ccgtgccggc ctacttcaac gactcgcagc
     1201 gccaggccac caaggatgcg ggtgtgatcg cggggctcaa cgtgctgcgg atcatcaacg
     1261 agcccacggc cgccgccatc gcctacggcc tggacagaac gggcaagggg gagcgcaacg
     1321 tgctcatctt tgacctgggc gggggcacct tcgacgtgtc catcctgacg atcgacgacg
     1381 gcatcttcga ggtgaaggcc acggccgggg acacccacct gggtggggag gactttgaca
     1441 acaggctggt gaaccacttc gtggaggagt tcaagagaaa acacaagaag gacatcagcc
     1501 agaacaagcg agccgtgagg cggctgcgca ccgcctgcga gagggccaag aggaccctgt
     1561 cgtccagcac ccaggccagc ctggagatcg actccctgtt tgagggcatc gacttctaca
     1621 cgtccatcac cagggcgagg ttcgaggagc tgtgctccga cctgttccga agcaccctgg
     1681 agcccgtgga gaaggctctg cgcgacgcca agctggacaa ggcccagatt cacgacctgg
     1741 tcctggtcgg gggctccacc cgcatcccca aggtgcagaa gctgctgcag gacttcttca
     1801 acgggcgcga cctgaacaag agcatcaacc ccgacgaggc tgtggcctac ggggcggcgg
     1861 tgcaggcggc catcctgatg ggggacaagt ccgagaacgt gcaggacctg ctgctgctgg
     1921 acgtggctcc cctgtcgctg gggctggaga cggccggagg cgtgatgact gccctgatca
     1981 agcgcaactc caccatcccc accaagcaga cgcagatctt caccacctac tccgacaacc
     2041 aacccggggt gctgatccag gtgtacgagg gcgagagggc catgacgaaa gacaacaatc
     2101 tgttggggcg cttcgagctg agcggcatcc ctccggcccc caggggcgtg ccccagatcg
     2161 aggtgacctt cgacatcgat gccaacggca tcctgaacgt cacggccacg gacaagagca
     2221 ccggcaaggc caacaagatc accatcacca acgacaaggg ccgcctgagc aaggaggaga
     2281 tcgagcgcat ggtgcaggag gcggagaagt acaaagcgga ggacgaggtg cagcgcgaga
     2341 gggtgtcagc caagaacgcc ctggagtcct acgccttcaa catgaagagc gccgtggagg
     2401 atgaggggct caagggcaag atcagcgagg ccgacaagaa gaaggtgctg gacaagtgtc
     2461 aagaggtcat ctcgtggctg gacgccaaca ccttggccga gaaggacgag tttgagcaca
     2521 agaggaagga gctggagcag gtgtgtaacc ccatcatcag cggactgtac cagggtgccg
     2581 gtggtcccgg gcctgggggc ttcggggctc agggtcccaa gggagggtct gggtcaggcc
     2641 ccaccattga ggaggtagat taggcggccg ctcgagtcta gagggcccgt ttaaacccgc
     2701 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     2761 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     2821 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     2881 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     2941 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     3001 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3061 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3121 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3181 gaccccaaaa aacttgatta gggtgatggt tcacgtacct agaagttcct attccgaagt
     3241 tcctattctc tagaaagtat aggaacttcc ttggccaaaa agcctgaact caccgcgacg
     3301 tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg
     3361 gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg
     3421 gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg
     3481 gccgcgctcc cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat
     3541 tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc
     3601 gctgttctgc agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag
     3661 acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat
     3721 ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc
     3781 gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc
     3841 gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc
     3901 cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc
     3961 gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc
     4021 gagcggaggc atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt
     4081 ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg
     4141 cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc
     4201 gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga
     4261 aaccgacgcc ccagcactcg tccgagggca aaggaatagc acgtactacg agatttcgat
     4321 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4381 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     4441 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4501 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     4561 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     4621 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4681 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4741 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4801 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4861 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4921 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4981 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5041 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5101 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5161 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5221 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5281 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5341 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5401 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5461 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5521 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5581 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5641 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5701 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5761 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5821 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5881 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5941 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6001 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6061 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6121 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6181 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6241 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6301 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6361 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6421 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6481 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6541 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6601 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6661 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6721 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     6781 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     6841 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     6901 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
     6961 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
     7021 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     7081 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     7141 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     7201 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     7261 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     7321 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     7381 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     7441 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     7501 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     7561 atataagcag agctctccct atcagtgata gagatctccc tatcagtgat agagatcgtc
     7621 gacgagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     7681 tccatagaag acaccgggac cgatccagcc tccggactct agcgtttaaa ctta
//
