LOCUS       Exported                7657 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA5/FRT/TO HSPH2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7657)
  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 7657)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7657
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     complement(3609..3626)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3615..3839
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3972..3991)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4123..4170
                     /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             4178..5200
                     /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    5330..5451
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5367..5386)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5421..5440
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5500..5516)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5500..5516)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5513..5535)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5524..5540
                     /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(5548..5578)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5593..5614
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5731..5748)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5902..6490)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5982..6001)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6661..7521)
                     /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     7284..7303
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7522..7626)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtaccg agctcggatc cactagtcca gtgtggtgga
     1021 attctgcaga tatcaccatg tcggtggtgg gcatagacct gggcttccag agctgctacg
     1081 tcgctgtggc ccgcgccggc ggcatcgaga ctatcgctaa tgagtatagc gaccgctgca
     1141 cgccggcttg catttctttt ggtcctaaga atcgttcaat tggagcagca gctaaaagcc
     1201 aggtaatttc taatgcaaag aacacagtcc aaggatttaa aagattccat ggccgagcat
     1261 tctctgatcc atttgtggag gcagaaaaat ctaaccttgc atatgatatt gtgcagttgc
     1321 ctacaggatt aacaggtata aaggtgacat atatggagga agagcgaaat tttaccactg
     1381 agcaagtgac tgccatgctt ttgtccaaac tgaaggagac agccgaaagt gttcttaaga
     1441 agcctgtagt tgactgtgtt gtttcggttc cttgtttcta tactgatgca gaaagacgat
     1501 cagtgatgga tgcaacacag attgctggtc ttaattgctt gcgattaatg aatgaaacca
     1561 ctgcagttgc tcttgcatat ggaatctata agcaggatct tcctgcctta gaagagaaac
     1621 caagaaatgt agtttttgta gacatgggcc actctgctta tcaagtttct gtatgtgcat
     1681 ttaatagagg aaaactgaaa gttctggcca ctgcatttga cacgacattg ggaggtagaa
     1741 aatttgatga agtgttagta aatcacttct gtgaagaatt tgggaagaaa tacaagctag
     1801 acattaagtc caaaatccgt gcattattac gactctctca ggagtgtgag aaactcaaga
     1861 aattgatgag tgcaaatgct tcagatctcc ctttgagcat tgaatgtttt atgaatgatg
     1921 ttgatgtatc tggaactatg aatagaggca aatttctgga gatgtgcaat gatctcttag
     1981 ctagagtgga gccaccactt cgtagtgttt tggaacaaac caagttaaag aaagaagata
     2041 tttatgcagt ggagatagtt ggtggtgcta cacgaatccc tgcggtaaaa gagaagatca
     2101 gcaaattttt cggtaaagaa cttagtacaa cattaaatgc tgatgaagct gtcactcgag
     2161 gctgtgcatt gcagtgtgcc atcttatcgc ctgctttcaa agtcagagaa ttttctatca
     2221 ctgatgtagt accatatcca atatctctga gatggaattc tccagctgaa gaagggtcaa
     2281 gtgactgtga agtcttttcc aaaaatcatg ctgctccttt ctctaaagtt cttacatttt
     2341 atagaaagga acctttcact cttgaggcct actacagctc tcctcaggat ttgccctatc
     2401 cagatcctgc tatagctcag ttttcagttc agaaagtcac tcctcagtct gatggctcca
     2461 gttcaaaagt gaaagtcaaa gttcgagtaa atgtccatgg cattttcagt gtgtccagtg
     2521 catctttagt ggaggttcac aagtctgagg aaaatgagga gccaatggaa acagatcaga
     2581 atgcaaagga ggaagagaag atgcaagtgg accaggagga accacatgtt gaagagcaac
     2641 agcagcagac accagcagaa aataaggcag agtctgaaga aatggagacc tctcaagctg
     2701 gatccaagga taaaaagatg gaccaaccac cccaagccaa gaaggcaaaa gtgaagacca
     2761 gtactgtgga cctgccaatc gagaatcagc tattatggca gatagacaga gagatgctca
     2821 acttgtacat tgaaaatgag ggtaagatga tcatgcagga taaactggag aaggagcgga
     2881 atgatgctaa gaacgcagtg gaggaatatg tgtatgaaat gagagacaag cttagtggtg
     2941 aatatgagaa gtttgtgagt gaagatgatc gtaacagttt tactttgaaa ctggaagata
     3001 ctgaaaattg gttgtatgag gatggagaag accagccaaa gcaagtttat gttgataagt
     3061 tggctgaatt aaaaaatcta ggtcaaccta ttaagatacg tttccaggaa tctgaagaac
     3121 gaccaaaatt atttgaagaa ctagggaaac agatccaaca gtatatgaaa ataatcagct
     3181 ctttcaaaaa caaggaggac cagtatgatc atttggatgc tgctgacatg acaaaggtag
     3241 aaaaaagcac aaatgaagca atggagtgga tgaataacaa gctaaatctg cagaacaagc
     3301 agagtttgac catggatcca gttgtcaagt caaaagagat tgaagctaaa attaaggagc
     3361 tgacaagtac ttgtagccct ataatttcaa agcccaaacc caaagtggaa cctccaaaag
     3421 aggaacaaaa aaatgcagag cagaatggac cagtggatgg acaaggagac aacccaggcc
     3481 cccaggctgc tgagcagggt acagacacag ctgtgccttc ggattcagac aagaagcttc
     3541 ctgaaatgga cattgattga ttccgcggcc gctcgagtct agagggcccg tttaaacccg
     3601 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3661 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3721 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3781 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     3841 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3901 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3961 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4021 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4081 cgaccccaaa aaacttgatt agggtgatgg ttcacgtacc tagaagttcc tattccgaag
     4141 ttcctattct ctagaaagta taggaacttc cttggccaaa aagcctgaac tcaccgcgac
     4201 gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc
     4261 ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg
     4321 ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc
     4381 ggccgcgctc ccgattccgg aagtgcttga cattggggaa ttcagcgaga gcctgaccta
     4441 ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc
     4501 cgctgttctg cagccggtcg cggaggccat ggatgcgatc gctgcggccg atcttagcca
     4561 gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga
     4621 tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac
     4681 cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc
     4741 cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg
     4801 ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt
     4861 cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt
     4921 cgagcggagg catccggagc ttgcaggatc gccgcggctc cgggcgtata tgctccgcat
     4981 tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc
     5041 gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat
     5101 cgcccgcaga agcgcggccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg
     5161 aaaccgacgc cccagcactc gtccgagggc aaaggaatag cacgtactac gagatttcga
     5221 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5281 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     5341 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5401 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5461 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5521 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5581 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5641 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5701 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5761 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5821 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5881 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5941 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6001 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6061 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6121 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6181 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6241 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6301 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6361 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6421 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6481 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6541 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6601 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6661 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6721 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6781 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6841 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6901 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6961 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7021 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7081 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7141 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7201 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7261 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7321 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7381 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7441 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7501 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7561 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7621 tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
