LOCUS       sequence-307704-        7217 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Heat shock chaperone 70 in Mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7217)
  AUTHORS   Zhou D, Li P, Lin Y, Lott JM, Hislop AD, Canaday DH, Brutkiewicz RR,
            Blum JS
  TITLE     Lamp-2a facilitates MHC class II presentation of cytoplasmic 
            antigens.
  JOURNAL   Immunity. 2005 May;22(5):571-81.
  PUBMED    15894275
REFERENCE   2  (bases 1 to 7217)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Immunity.";
            date: "2005-05"; volume: "22(5)"; pages: "571-81"
COMMENT     Sequence Label: pCDNAZeo(-)HSC73
FEATURES             Location/Qualifiers
     source          1..7217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        131..510
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        511..714
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     665..685
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     759..778
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        759..777
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3121..3138)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3127..3351
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3397..3825
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3484..3503)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3694..3715
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3834..3854)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3839..4168
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4019..4154
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4081..4100
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4216..4263
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4282..4656
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4786..4907
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4823..4842)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4877..4896
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4956..4972)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4956..4972)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4969..4991)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4980..4996
                     /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(join(5004..5010,5011..5028,5029..5034))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5049..5070
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5187..5204)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5358..5946)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5438..5457)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6117..6908,6909..6977))
                     /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     6740..6759
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6978..7082)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7157..7176)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
       61 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
      121 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
      181 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      241 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      301 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      361 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      421 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      481 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      541 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      601 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      661 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg
      721 ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag
      781 acccaagctg gctagcgttt aaacgggccc tctagactcg agcggccgcc actgtgctgg
      841 atatctgcag aattccacca cactggacta gtggatccat gtccaaggga cctgcagttg
      901 gtattgatct tggcaccacc tactcttgtg tgggtgtttt ccagcacgga aaagtcgaga
      961 taattgccaa tgatcaggga aaccgaacca ctccaagcta tgtcgccttt acggacactg
     1021 aacggttgat cggtgatgcc gcaaagaatc aagttgcaat gaaccccacc aacacagttt
     1081 ttgatgccaa acgtctgatt ggacgcagat ttgatgatgc tgttgtccag tctgatatga
     1141 aacattggcc ctttatggtg gtgaatgatg ctggcaggcc caaggtccaa gtagaataca
     1201 agggagagac caaaagcttc tatccagagg aggtgtcttc tatggttctg acaaagatga
     1261 aggaaattgc agaagcctac cttgggaaga ctgttaccaa tgctgtggtc acagtgccag
     1321 cttactttaa tgactctcag cgtcaggcta ccaaagatgc tggaactatt gctggtctca
     1381 atgtacttag aattattaat gagccaactg ctgctgctat tgcttacggc ttagacaaaa
     1441 aggttggagc agaaagaaac gtgctcatct ttgacctggg aggtggcact tttgatgtgt
     1501 caatcctcac tattgaggat ggaatctttg aggtcaagtc tacagctgga gacacccact
     1561 tgggtggaga agattttgac aaccgaatgg tcaaccattt tattgctgag tttaagcgca
     1621 agcataagaa ggacatcagt gagaacaaga gagctgtaag acgcctccgt actgcttgtg
     1681 aacgtgctaa gcgtaccctc tcttccagca cccaggccag tattgagatc gattctctct
     1741 atgaaggaat cgacttctat acctccatta cccgtgcccg atttgaagaa ctgaatgctg
     1801 acctgttccg tggcaccctg gacccagtag agaaagccct tcgagatgcc aaactagaca
     1861 agtcacagat tcatgatatt gtcctggttg gtggttctac tcgtatcccc aagattcaga
     1921 agcttctcca agacttcttc aatggaaaag aactgaataa gagcatcaac cctgatgaag
     1981 ctgttgctta tggtgcagct gtccaggcag ccatcttgtc tggagacaag tctgagaatg
     2041 ttcaagattt gctgctctcg gatgtcactc ctctttccct tggtattgaa actgctggtg
     2101 gagtcatgac tgtcctcatc aagcgtaata ccaccattcc taccaagcag acacagacct
     2161 tcactaccta ttctgacaac cagcctggtg tgcttattca ggtttatgaa ggcgagcgtg
     2221 ccatgacaaa ggataacaac ctgcttggca agtttgaact cacaggcata cctcctgcac
     2281 cccgaggtgt tcctcagatt gaagtcactt ttgacattga tgccaatggt atcctcaatg
     2341 tctctgctgt ggacaagagt acgggaaaag agaacaagat tactatcact aatgacaagg
     2401 gccgtttgag caaggaagac attgaacgta tggtccagga agctgagaag tacaaagctg
     2461 aagatgagaa gcagagggac aaggtgtcat ccaagaattc acttgagtcc tatgccttca
     2521 acatgaaagc aactgttgaa gatgagaaac ttcaaggcaa gattaacgat gaggacaaac
     2581 agaagattct ggacaagtgt aatgaaatta tcaactggct tgataagaat cagactgctg
     2641 agaaggaaga atttgaacat caacagaaag agctggagaa agtttgcaac cccatcatca
     2701 ccaagctgta ccagagtgca ggaggcatgc caggaggaat gcctggggga tttcctggtg
     2761 gtggagctcc tccctctggt ggtgcttcct cagggcccac cattgaagag gttgattaag
     2821 ccaaccaagt gtagatgtag cattgttcca cacatttaaa acatttgaag gacctaaatt
     2881 cgtagcaaat tctgtggcag ttttaaaaag ttaagctgct atagtaagtt actgggcatt
     2941 ctcaatactt gaatatggaa catatgcaca ggggaaggaa ataacattgc actttataaa
     3001 cactgtattg taagtggaaa atgcaatgtc ttaaataaaa ctatttaaaa ttggcacgga
     3061 attcctgcag cccgggggat ccgagctcgg taccaagctt aagtttaaac cgctgatcag
     3121 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     3181 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     3241 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     3301 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     3361 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     3421 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3481 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3541 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3601 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     3661 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     3721 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     3781 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     3841 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3901 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3961 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4021 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4081 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     4141 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     4201 atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac
     4261 gacaaggtga ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg
     4321 cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc
     4381 gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc
     4441 caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag
     4501 ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc
     4561 atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc
     4621 aactgcgtgc acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc
     4681 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4741 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     4801 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4861 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     4921 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4981 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5041 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5101 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5161 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5221 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5281 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5341 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5401 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5461 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5521 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5581 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5641 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5701 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5761 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5821 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5881 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5941 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6001 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6061 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6121 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6181 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6241 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6301 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6361 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6421 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6481 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6541 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6601 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6661 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6721 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6781 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6841 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6901 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6961 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7021 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7081 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     7141 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     7201 ctgcttgtgt gttggag
//
