LOCUS       IsHSP70mCherry        6002 bp DNA     circular SYN 16-APR-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6002)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 16, 2024 from SnapGene 7.2.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6002
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     misc_feature    26..1326
                     /label=HS70 Promoter
     misc_feature    874..1326
                     /label=Portion Used on Is3xP3
     misc_feature    879..885
                     /label=TATA Box (TATAWAW)
     primer_bind     1374..1400
                     /label=mCherryBglIIkozakstartFor
     regulatory      1374..1379
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1380..2129
                     /codon_start=1
                     /label=mCherry
                     /translation="MGPGLGMLSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGE
                     GRPYEGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKW
                     ERVMNFEDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYP
                     EDGALKGEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIV
                     EQYERAEGRHSTGGMDELYKGLEGGAP"
     misc_feature    1385..2125
                     /label=mCherry(no SbfI)
     primer_bind     complement(2115..2134)
                     /label=mCherry MFE1 STOP reverse primer
     misc_feature    2276
                     /label=mfeI
     polyA_signal    2290..2411
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        2514..2871
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2722..2857
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2902..3696
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3721..3769
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4096..4684)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(4884..5675,5676..5744))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    5849..5897
                     /note="synthetic polyadenylation signal"
     misc_feature    5911..6002
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5951..5970
                     /label=RV3 Primer
ORIGIN
        1 ggcctaactg gccggtacct gagctcgccc agtttaaggt gtgtgcgttt ggaatatgtg
       61 tagtacacag aaccttagat cacgcttaca ttgaacaata aagtggcaga ttaaccttta
      121 agtgaatccc cccaattcgc accaaaaaaa aaaaaatgct gggttaatca aacagctggt
      181 tttctattta ttcagtggaa aatccaaagg aaggttttac cttccgttaa gatccgtttg
      241 gtatggtggc caatgtcaac tgcccgtatt caacacaatg ttatgcaaca gaagttggaa
      301 cctgcggaac atttccacat cttcacaagt tccgtgatgg cgcaaggtgg ataacgcgat
      361 ttctctgaaa ggtgtgggcg gctcccaaaa tgctcgaaag gagccgcgaa gacgcccact
      421 acccccgcct tcgcattgtg gcgtgaagtg tagtggcaag aaacctagga aaagtctcga
      481 gcatactggg tgtggcggcc tgaacatgtg gccagcaaag tcaaaatttt aaacgcacgt
      541 tattttcagg ataatgctta cctccattca taacattcct ttgtttttgt gtgtgtctgt
      601 gtgcgcgcgc actttatgtt ttatttcaag aaaagaaaca aaaaagaaga aaaaaaaaaa
      661 aacagcgagc gatcaagaaa caagacgact tgtgttgtcg cccattgaat actttttgtt
      721 ttcttctagg atgagaattt ttagtctttg gcatgttccc cgctcaattt tccgtacatg
      781 tgcttgtcac taactcgacg cgcatttttc gtttttcttt tttgtttttt attctttttc
      841 gcgtgtgttt ttgtttaact tttttaattt tttctttgta taaaataaag gtcattctct
      901 cattcaacga gggatcacgt tttcgaatgt tccagaagta aaggtacccg cgtgcttcca
      961 acaaacagca aaccaaaaat agcgcggccg aaccagaaga gagcagacga cgcgaggaga
     1021 ttctggcgct ttcgattgcg caacagccaa tagcaagcgc tctctgaata cgtcacgcgg
     1081 aggctcgctc ctgattggat ggggcgcctg agcaatttct cgaagcttct ggaagggaga
     1141 gcggagcgta caaaaaggcg ggccgcgctc ttcccgccca tacgttgctc cgagagcgca
     1201 acttttgaaa actgatcgac ctagcgccga gtgagacgac agaaacccgg aaaacaacca
     1261 tgccaatccg aagcaagagt ggagcgccgg ccatcggcat tgaccttggc acaacctact
     1321 cgtgcagatc tggcctcggc ggccaagctt ggcaatccgg tactgttggt aaagccacca
     1381 tgggcccggg cctagggatg ttgagcaagg gcgaggagga taacatggcc atcatcaagg
     1441 agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg
     1501 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
     1561 agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca
     1621 aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg
     1681 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg
     1741 actcctccct acaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc
     1801 cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga
     1861 tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg
     1921 gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc
     1981 ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca
     2041 tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt
     2101 acaagggtct agaggggggc gcgccttaag aattctgcag tcgacggtac cgcgggcccg
     2161 ggatccaccg gatctagata actgatcata atcagccata ccacatttgt agaggtttta
     2221 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     2281 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     2341 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     2401 aatgtatctt aaagccctta ggggatcctt tgcgtattgg gcgctcttcc gctgatctgc
     2461 gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttagctac cttctgaggc
     2521 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     2581 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     2641 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     2701 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     2761 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     2821 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcgat
     2881 tcttctgaca ctagcgccac catgatcgaa caagacggcc tccatgctgg cagtcccgca
     2941 gcttgggtcg aacgcttgtt cgggtacgac tgggcccagc agaccatcgg atgtagcgat
     3001 gcggccgtgt tccgtctaag cgctcaaggc cggcccgtgc tgttcgtgaa gaccgacctg
     3061 agcggcgccc tgaacgagct tcaagacgag gctgcccgcc tgagctggct ggccaccacc
     3121 ggcgtaccct gcgccgctgt gttggatgtt gtgaccgaag ccggccggga ctggctgctg
     3181 ctgggcgagg tccctggcca ggatctgctg agcagccacc ttgcccccgc tgagaaggtt
     3241 tctatcatgg ccgatgcaat gcggcgcctg cacaccctgg accccgctac ctgccccttc
     3301 gaccaccagg ctaagcatcg gatcgagcgt gctcggaccc gcatggaggc cggcctggtg
     3361 gaccaggacg acctggacga ggagcatcag ggcctggccc ccgctgaact gttcgcccga
     3421 ctgaaagccc gcatgccgga cggtgaggac ctggttgtca cacacggaga tgcctgcctc
     3481 cctaacatca tggtcgagaa tggccgcttc tccggcttca tcgactgcgg tcgcctagga
     3541 gttgccgacc gctaccagga catcgccctg gccacccgcg acatcgctga ggagcttggc
     3601 ggcgagtggg ccgaccgctt cttagtcttg tacggcatcg cagctcccga cagccagcgc
     3661 atcgccttct accgcttgct cgacgagttc ttttaatgat ctagaaccgg tcatggccgc
     3721 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtgt tcgaactaga
     3781 tgctgtcgac cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
     3841 ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
     3901 gtgccggcag cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3961 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4021 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4081 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4141 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4201 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4261 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4321 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4381 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4441 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4501 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4561 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4621 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4681 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4741 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4801 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagcggccg
     4861 caaatgctaa accactgcag tggttaccag tgcttgatca gtgaggcacc gatctcagcg
     4921 atctgcctat ttcgttcgtc catagtggcc tgactccccg tcgtgtagat cactacgatt
     4981 cgtgagggct taccatcagg ccccagcgca gcaatgatgc cgcgagagcc gcgttcaccg
     5041 gcccccgatt tgtcagcaat gaaccagcca gcagggaggg ccgagcgaag aagtggtcct
     5101 gctactttgt ccgcctccat ccagtctatg agctgctgtc gtgatgctag agtaagaagt
     5161 tcgccagtga gtagtttccg aagagttgtg gccattgcta ctggcatcgt ggtatcacgc
     5221 tcgtcgttcg gtatggcttc gttcaactct ggttcccagc ggtcaagccg ggtcacatga
     5281 tcacccatat tatgaagaaa tgcagtcagc tccttagggc ctccgatcgt tgtcagaagt
     5341 aagttggccg cggtgttgtc gctcatggta atggcagcac tacacaattc tcttaccgtc
     5401 atgccatccg taagatgctt ttccgtgacc ggcgagtact caaccaagtc gttttgtgag
     5461 tagtgtatac ggcgaccaag ctgctcttgc ccggcgtcta tacgggacaa caccgcgcca
     5521 catagcagta ctttgaaagt gctcatcatc gggaatcgtt cttcggggcg gaaagactca
     5581 aggatcttgc cgctattgag atccagttcg atatagccca ctcttgcacc cagttgatct
     5641 tcagcatctt ttactttcac cagcgtttcg gggtgtgcaa aaacaggcaa gcaaaatgcc
     5701 gcaaagaagg gaatgagtgc gacacgaaaa tgttggatgc tcatactcgt cctttttcaa
     5761 tattattgaa gcatttatca gggttactag tacgtctctc aaggataagt aagtaatatt
     5821 aaggtacggg aggtattgga caggccgcaa taaaatatct ttattttcat tacatctgtg
     5881 tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa acaaaacgaa
     5941 acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct
     6001 ct
//
