LOCUS       sequence-345198-        3516 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HSP70B promoter for fusion with bacterial promoters.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3516)
  AUTHORS   Matinyan N, Karkhanis MS, Gonzalez Y, Jain A, Saltzman A, 
            Malovannaya A, Sarrion-Perdigones A, Dierick HA, Venken KJT
  TITLE     Multiplexed drug-based selection and counterselection genetic 
            manipulations in Drosophila.
  JOURNAL   Cell Rep. 2021 Sep 14;36(11):109700. doi: 
            10.1016/j.celrep.2021.109700.
  PUBMED    34525356
REFERENCE   2  (bases 1 to 3516)
  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: "Cell Rep.";
            date: "2021-09-14"; volume: "36(11):109700. doi"; pages: " 
            10.1016/j.celrep.2021.109700"
COMMENT     Sequence Label: pDmHsp70b-TACT
FEATURES             Location/Qualifiers
     source          1..3516
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     132..151
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     385..402
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    519..540
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(555..560,561..578,579..585)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    593..609
                     /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)."
     primer_bind     598..620
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     617..633
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     617..633
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        651..669
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     651..668
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(775..1226)
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     complement(806..827)
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     regulatory      1264..1273
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1268..1277
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1307..1326)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1308..1326)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1333..1350)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1333..1349)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1342..1364)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1490..1945
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1577..1596)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1787..1808
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2023..2127
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2128..2196,2197..2988)
                     /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     complement(2346..2365)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3159..3516,1..231)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
       61 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      121 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      181 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      241 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      301 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
      361 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
      421 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
      481 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
      541 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
      601 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct atttaggtga
      661 cactatagaa tactcaagct atgcatccaa cgcgttggga gctctcccat atggtcgacc
      721 tgcaggcggc cgcgaattca ctagtgattg ggcgatgagt ggtctcgcat tttgagagtt
      781 ctcttcttgt attcaataat tacttcttgg cagatttcag tagttgcagt tgatttactt
      841 ggttgctggt tacttttaat tgattcactt taacttgcac tttactgcag attgtttagc
      901 ttgttcagct gcgcttgttt atttgcttag ctttcgctta gcgacgtgtt cactttgctt
      961 gtttgaattg aattgtcgct ccgtagacga agcgcctcta tttatactcc ggcgctcttt
     1021 tcgcgaacat tcgaggcgcg ctctctcgaa ccaacgagag cagtatgccg tttactgtgt
     1081 gacagagtga gagagcatta gtgcagagag ggagcgaccc aaaaagaaaa gagagaataa
     1141 cgaataacgg ccagagaaat ttctcgagtt ttctttctgc caaacaaatg acctaccaca
     1201 ataaccagtt tgttttggga ttctagctcc cgagaccact catcgcccaa tcgaattccc
     1261 gcggccgcca tggcggccgg gagcatgcga cgtcgggccc aattcgccct atagtgagtc
     1321 gtattacaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     1381 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     1441 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga cgcgccctgt
     1501 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     1561 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     1621 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     1681 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc agcgccctga
     1741 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     1801 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     1861 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     1921 aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg
     1981 gtatttcaca ccgcatcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     2041 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     2101 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     2161 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     2221 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     2281 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     2341 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     2401 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     2461 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     2521 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     2581 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     2641 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     2701 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     2761 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     2821 aaatctggag ccggtgagcg tgggtcgcgc ggtatcattg cagcactggg gccagatggt
     2881 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     2941 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     3001 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     3061 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     3121 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     3181 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     3241 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     3301 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     3361 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     3421 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     3481 gggggttcgt gcacacagcc cagcttggag cgaacg
//
