LOCUS       Exported                7104 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA5/FRT/TO V5 HSPA6
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7104)
  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 7104)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7104
                     /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"
     CDS             996..1037
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3056..3073)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3062..3286
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3419..3438)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3570..3617
                     /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             3625..4647
                     /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    4777..4898
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4814..4833)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4868..4887
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4947..4963)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4947..4963)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4960..4982)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4971..4987
                     /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(4995..5025)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5040..5061
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5178..5195)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5349..5937)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5429..5448)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6108..6968)
                     /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     6731..6750
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6969..7073)
                     /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 cttgaattca ccatgggtaa gcctatccct aaccctctcc
     1021 tcggtctcga ttctacggga tccactagtc cagtgtggtg gaattctgca gatatcacca
     1081 tgcaggcccc acgggagctc gcggtgggca tcgacctggg caccacctac tcgtgcgtgg
     1141 gcgtgtttca gcagggccgc gtggagatcc tggccaacga ccagggcaac cgcaccacgc
     1201 ccagctacgt ggccttcacc gacaccgagc ggctggtcgg ggacgcggcc aagagccagg
     1261 cggccctgaa cccccacaac accgtgttcg atgccaagcg gctgatcggg cgcaagttcg
     1321 cggacaccac ggtgcagtcg gacatgaagc actggccctt ccgggtggtg agcgagggcg
     1381 gcaagcccaa ggtgcgcgta tgctaccgcg gggaggacaa gacgttctac cccgaggaga
     1441 tctcgtccat ggtgctgagc aagatgaagg agacggccga ggcgtacctg ggccagcccg
     1501 tgaagcacgc agtgatcacc gtgcccgcct atttcaatga ctcgcagcgc caggccacca
     1561 aggacgcggg ggccatcgcg gggctcaacg tgttgcggat catcaatgag cccacggcag
     1621 ctgccatcgc ctatgggctg gaccggcggg gcgcgggaga gcgcaacgtg ctcatttttg
     1681 acctgggtgg gggcaccttc gatgtgtcgg ttctctccat tgacgctggt gtctttgagg
     1741 tgaaagccac tgctggagat acccacctgg gaggagagga cttcgacaac cggctcgtga
     1801 accacttcat ggaagaattc cggcggaagc atgggaagga cctgagcggg aacaagcgtg
     1861 ccctgcgcag gctgcgcaca gcctgtgagc gcgccaagcg caccctgtcc tccagcaccc
     1921 aggccaccct ggagatagac tccctgttcg agggcgtgga cttctacacg tccatcactc
     1981 gtgcccgctt tgaggaactg tgctcagacc tcttccgcag caccctggag ccggtggaga
     2041 aggccctgcg ggatgccaag ctggacaagg cccagattca tgacgtcgtc ctggtggggg
     2101 gctccactcg catccccaag gtgcagaagt tgctgcagga cttcttcaac ggcaaggagc
     2161 tgaacaagag catcaaccct gatgaggctg tggcctatgg ggctgctgtg caggcggccg
     2221 tgttgatggg ggacaaatgt gagaaagtgc aggatctcct gctgctggat gtggctcccc
     2281 tgtctctggg gctggagaca gcaggtgggg tgatgaccac gctgatccag aggaacgcca
     2341 ctatccccac caagcagacc cagactttca ccacctactc ggacaaccag cctggggtct
     2401 tcatccaggt gtatgagggt gagagggcca tgaccaagga caacaacctg ctggggcgtt
     2461 ttgaactcag tggcatccct cctgccccac gtggagtccc ccagatagag gtgacctttg
     2521 acattgatgc taatggcatc ctgagcgtga cagccactga caggagcaca ggtaaggcta
     2581 acaagatcac catcaccaat gacaagggcc ggctgagcaa ggaggaggtg gagaggatgg
     2641 ttcatgaagc cgagcagtac aaggctgagg atgaggccca gagggacaga gtggctgcca
     2701 aaaactcgct ggaggcccat gtcttccatg tgaaaggttc tttgcaagag gaaagcctta
     2761 gggacaagat tcccgaagag gacaggcgca aaatgcaaga caagtgtcgg gaagtccttg
     2821 cctggctgga gcacaaccag ctggcagaga aggaggagta tgagcatcag aagagggagc
     2881 tggagcaaat ctgtcgcccc atcttctcca ggctctatgg ggggcctggt gtccctgggg
     2941 gcagcagttg tggcactcaa gcccgccagg gggaccccag caccggcccc atcattgagg
     3001 aggttgattg agcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg
     3061 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3121 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3181 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     3241 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     3301 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3361 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3421 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3481 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3541 cttgattagg gtgatggttc acgtacctag aagttcctat tccgaagttc ctattctcta
     3601 gaaagtatag gaacttcctt ggccaaaaag cctgaactca ccgcgacgtc tgtcgagaag
     3661 tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa
     3721 tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc
     3781 gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg
     3841 attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc
     3901 cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag
     3961 ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc
     4021 ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg
     4081 attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc
     4141 gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac
     4201 ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg
     4261 gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc
     4321 ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat
     4381 ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa
     4441 ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc
     4501 gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc
     4561 gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc
     4621 agcactcgtc cgagggcaaa ggaatagcac gtactacgag atttcgattc caccgccgcc
     4681 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4741 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     4801 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4861 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     4921 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4981 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     5041 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5101 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5161 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5221 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5281 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5341 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5401 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5461 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5521 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5581 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5641 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5701 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5761 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5821 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5881 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5941 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6001 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6061 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6121 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6181 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6241 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6301 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6361 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6421 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6481 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6541 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6601 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6661 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6721 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6781 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6841 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6901 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6961 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7021 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7081 ccccgaaaag tgccacctga cgtc
//
