LOCUS       sequence-485413-        4293 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SmBit-CHIP expression with K30A substitution.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4293)
  AUTHORS   Stewart M, Paththamperuma C, McCann C, Cottingim K, Zhang H, 
            DelVecchio R, Peng I, Fennimore E, Nix JC, Saeed MN, George K, 
            Makaroff M, Colie M, Paulakonis E, Almeida MF, Afolayan AJ, Brown 
            NG, Page RC, Schisler JC
  TITLE     Phosphorylation-State Modulated Binding of HSP70: Structural 
            Insights and Compensatory Protein Engineering
  JOURNAL   bioRxiv 2025.02.17.637997
REFERENCE   2  (bases 1 to 4293)
  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: "bioRxiv 
            2025.02.17.637997"
COMMENT     Sequence Label: pSmBit-CHIP-K30A
FEATURES             Location/Qualifiers
     source          1..4293
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        28..779
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      809..818
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1847..1968
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1858..1877)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1912..1931
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2216..2233)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2387..2975)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2467..2486)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3175..3966,3967..4035))
                     /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    4140..4188
                     /note="synthetic polyadenylation signal"
     misc_feature    4202..4293
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4242..4261
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagtctaaat gagtcttcgg acctcgcggg ggccgcttaa
       61 gcggtggtta gggtttgtct gacgcggggg gagggggaag gaacgaaaca ctctcattcg
      121 gaggcggctc ggggtttggt cttggtggcc acgggcacgc agaagagcgc cgcgatcctc
      181 ttaagcaccc ccccgccctc cgtggaggcg ggggtttggt cggcgggtgg taactggcgg
      241 gccgctgact cgggcgggtc gcgcgcccca gagtgtgacc ttttcggtct gctcgcagac
      301 ccccgggcgg cgccgccgcg gcggcgacgg gctcgctggg tcctaggctc catggggacc
      361 gtatacgtgg acaggctctg gagcatccgc acgactgcgg tgatattacc ggagaccttc
      421 tgcgggacga gccgggtcac gcggctgacg cggagcgtcc gttgggcgac aaacaccagg
      481 acggggcaca ggtacactat cttgtcaccc ggaggcgcga gggactgcag gagcttcagg
      541 gagtggcgca gctgcttcat ccccgtggcc cgttgctcgc gtttgctggc ggtgtccccg
      601 gaagaaatat atttgcatgt ctttagttct atgatgacac aaaccccgcc cagcgtcttg
      661 tcattggcga agtcgaacac gcagatgcag tcggggcggc gcggtcccag gtccacttcg
      721 catattaagg tgacgcgtgt ggcctcgaac accgagcgac cctgcagcga cccgcttaaa
      781 agcttggcaa tccggtactg ttggtaaagc caccatggtg accggctacc ggctgttcga
      841 ggagattctc gggagttccg gtggtggcgg gagcggaggt ggaggctcga gcggtatgaa
      901 gggcaaggag gagaaggagg gcggcgcacg gctgggcgct ggcggcggaa gccccgagaa
      961 gagcccgagc gcgcaggagc tcgcggagca gggcaatcgt ctgttcgtgg gccgaaagta
     1021 cccggaggcg gcggcctgct acggccgcgc gatcacccgg aacccgctgg tggccgtgta
     1081 ttacaccaac cgggccttgt gctacctgaa gatgcagcag cacgagcagg ccctggccga
     1141 ctgccggcgc gccctggagc tggacgggca gtctgtgaag gcgcacttct tcctggggca
     1201 gtgccagctg gagatggaga gctatgatga ggccatcgcc aatctgcagc gagcttacag
     1261 cctggccaag gagcagcggc tgaacttcgg ggacgacatc cccagcgctc ttcgaatcgc
     1321 gaagaagaag cgctggaaca gcattgagga gcggcgcatc caccaggaga gcgagctgca
     1381 ctcctacctc tccaggctca ttgccgcgga gcgtgagagg gagctggaag agtgccagcg
     1441 aaaccacgag ggtgatgagg acgacagcca cgtccgggcc cagcaggcct gcattgaggc
     1501 caagcacgac aagtacatgg cggacatgga cgagcttttt tctcaggtgg atgagaagag
     1561 gaagaagcga gacatccccg actacctgtg tggcaagatc agctttgagc tgatgcggga
     1621 gccgtgcatc acgcccagtg gcatcaccta cgaccgcaag gacatcgagg agcacctgca
     1681 gcgtgtgggt cattttgacc ccgtgacccg gagccccctg acccaggaac agctcatccc
     1741 caacttggct atgaaggagg ttattgacgc attcatctct gagaatggct gggtggagga
     1801 ctactgatct agagtcgggg cggccggccg cttcgagcag acatgataag atacattgat
     1861 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     1921 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     1981 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     2041 aacctctaca aatgtggtaa aatcgataag gatccgtcga ccgatgccct tgagagcctt
     2101 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
     2161 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc
     2221 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2281 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2341 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2401 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2461 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2521 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2581 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2641 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2701 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2761 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2821 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2881 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2941 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3001 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3061 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3121 tatatgagta aacttggtct gacagcggcc gcaaatgcta aaccactgca gtggttacca
     3181 gtgcttgatc agtgaggcac cgatctcagc gatctgccta tttcgttcgt ccatagtggc
     3241 ctgactcccc gtcgtgtaga tcactacgat tcgtgagggc ttaccatcag gccccagcgc
     3301 agcaatgatg ccgcgagagc cgcgttcacc ggcccccgat ttgtcagcaa tgaaccagcc
     3361 agcagggagg gccgagcgaa gaagtggtcc tgctactttg tccgcctcca tccagtctat
     3421 gagctgctgt cgtgatgcta gagtaagaag ttcgccagtg agtagtttcc gaagagttgt
     3481 ggccattgct actggcatcg tggtatcacg ctcgtcgttc ggtatggctt cgttcaactc
     3541 tggttcccag cggtcaagcc gggtcacatg atcacccata ttatgaagaa atgcagtcag
     3601 ctccttaggg cctccgatcg ttgtcagaag taagttggcc gcggtgttgt cgctcatggt
     3661 aatggcagca ctacacaatt ctcttaccgt catgccatcc gtaagatgct tttccgtgac
     3721 cggcgagtac tcaaccaagt cgttttgtga gtagtgtata cggcgaccaa gctgctcttg
     3781 cccggcgtct atacgggaca acaccgcgcc acatagcagt actttgaaag tgctcatcat
     3841 cgggaatcgt tcttcggggc ggaaagactc aaggatcttg ccgctattga gatccagttc
     3901 gatatagccc actcttgcac ccagttgatc ttcagcatct tttactttca ccagcgtttc
     3961 ggggtgtgca aaaacaggca agcaaaatgc cgcaaagaag ggaatgagtg cgacacgaaa
     4021 atgttggatg ctcatactcg tcctttttca atattattga agcatttatc agggttacta
     4081 gtacgtctct caaggataag taagtaatat taaggtacgg gaggtattgg acaggccgca
     4141 ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac
     4201 taacatacgc tctccatcaa aacaaaacga aacaaaacaa actagcaaaa taggctgtcc
     4261 ccagtgcaag tgcaggtgcc agaacatttc tct
//
