LOCUS       Exported                7658 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 7658)
  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 7658)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7658
                     /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(3610..3627)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3616..3840
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3973..3992)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4124..4171
                     /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             4179..5201
                     /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    5331..5452
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5368..5387)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5422..5441
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5501..5517)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5501..5517)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5514..5536)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5525..5541
                     /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(5549..5579)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5594..5615
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5732..5749)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5903..6491)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5983..6002)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6662..7522)
                     /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     7285..7304
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7523..7627)
                     /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 agtgctagta 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 gcagtgtacc 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 gagaaccagc 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 ggccaaccta ttaagatacg tttccaggaa tctgaagaac
     3121 gaccaaaatt atttgaagaa ctagggaaac agatccaaca gtatatgaaa ataatcagct
     3181 ctttcaaaaa caaggaggac cagtatgatc atttggatgc tgccgacatg gcaaaggtag
     3241 aaaaaagcac aaatgaagca atggagtgga tgaataacaa gctaaatctg cagaacaagc
     3301 agagtttgac catggatcca gttgtcaagt caaaagagat tgaagctaaa attaaggagc
     3361 tgacaagtac ttgtagccct ataatttcaa agcccaaacc caaagtggaa cctccaaaag
     3421 aggaacaaaa aaaatgcaga gcagaatgga ccagtggatg gacaaggaga caacccaggc
     3481 ccccaggctg ctgagcaggg tacagacaca gctgtgcctt cggattcaga caagaagctt
     3541 cctgaaatgg acattgattg attccgcggc cgctcgagtc tagagggccc gtttaaaccc
     3601 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     3661 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     3721 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     3781 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     3841 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     3901 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3961 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4021 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4081 tcgaccccaa aaaacttgat tagggtgatg gttcacgtac ctagaagttc ctattccgaa
     4141 gttcctattc tctagaaagt ataggaactt ccttggccaa aaagcctgaa ctcaccgcga
     4201 cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct
     4261 cggagggcga agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc
     4321 gggtaaatag ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat
     4381 cggccgcgct cccgattccg gaagtgcttg acattgggga attcagcgag agcctgacct
     4441 attgcatctc ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc
     4501 ccgctgttct gcagccggtc gcggaggcca tggatgcgat cgctgcggcc gatcttagcc
     4561 agacgagcgg gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg
     4621 atttcatatg cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca
     4681 ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc
     4741 ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg
     4801 gccgcataac agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg
     4861 tcgccaacat cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact
     4921 tcgagcggag gcatccggag cttgcaggat cgccgcggct ccgggcgtat atgctccgca
     4981 ttggtcttga ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg
     5041 cgcagggtcg atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa
     5101 tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt agaagtactc gccgatagtg
     5161 gaaaccgacg ccccagcact cgtccgaggg caaaggaata gcacgtacta cgagatttcg
     5221 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5281 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5341 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5401 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5461 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5521 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5581 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5641 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5701 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5761 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5821 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5881 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5941 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6001 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6061 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6121 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6181 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6241 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6301 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6361 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6421 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6481 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6541 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6601 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6661 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6721 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6781 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6841 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6901 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6961 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7021 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7081 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7141 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7201 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7261 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7321 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7381 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7441 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7501 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7561 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7621 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
