LOCUS       sequence-137518-        5529 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Allows expression of rabbit alpha-skeletal tropomyosin from a 
            rhamnose inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5529)
  AUTHORS   Eastwood T, Baker K, Brooker H, Frank S, Mulvihill DP
  TITLE     An enhanced recombinant amino-terminal acetylation system and novel 
            in vivo high-throughput screen for molecules affecting 
            alpha-synuclein oligomerisation.
  JOURNAL   FEBS Lett. 2017 Feb 18. doi: 10.1002/1873-3468.12597.
  PUBMED    28214355
REFERENCE   2  (bases 1 to 5529)
  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: "FEBS Lett. 
            2017 Feb 18. doi: 10.1002/1873-3468.12597."
COMMENT     Sequence Label: pETpRha-alpha-SkTm
FEATURES             Location/Qualifiers
     source          1..5529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..15,16..32,33..38)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(404..451)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     447..465
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     RBS             1378..1400
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(1418..1536)
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     primer_bind     complement(1604..1623)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             3081..3272
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3266..3288)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3374..3516
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3429..3448
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3531..3548)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3702..4290)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3782..3801)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4461..5252,5253..5321))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5084..5103
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5322..5426)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5494..5512
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
       61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
      121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
      181 gcgggatatc gtccattccg acagcatcgc cagtcactat ggcgtgctgc tagcgctata
      241 tgcgttgatg caatttctat gcgcacccgt tctcggagca ctgtccgacc gctttggccg
      301 ccgcccagtc ctgctcgctt cgctacttgg agccactatc gactacgcga tcatggcgac
      361 cacacccgtc ctgtggatat ccggatatag ttcctccttt cagcaaaaaa cccctcaaga
      421 cccgtttaga ggccccaagg ggttatgcta gttattgctc agcggtggca gcagccaact
      481 cagcttcctt tcgggctttg ttagcagccg gatccttata tggaagtcat atcgttgaga
      541 gcgtggtcca gctcctcgct gatggccttg tacttcagtt tctgagcata cagctcgtct
      601 tctaagtcat caatgctttt ctccaatttg gttactgatc tctctgcaaa ctcagcccgg
      661 gtctcagcct ccttcagctt gtcagagaga accttgatct cctcttcata cttgtcttct
      721 ttctgagagt acttctcagc ctgagcctcc agtgacttca agttgttcgt caccgttttc
      781 aactcttctt caagctcggc acatttgcct tccgagagct cagccctctc ctccgcacgc
      841 tccagatcgc tctcgatgat gaccagctta cgggccacct cttcatactt tcggtcagca
      901 tcttcagcaa tgtgcttggc ctctttcagc tggatctcct gaatctccat cttttcttca
      961 tctttttggg ctcggctttc aatgactttc atgcctctct cactctcatc tgcagccttc
     1021 tcagcctcct ccagcttctg tagagctgtg gccagacgct cctgagcgcg atccaactcc
     1081 tcctcaacca gctggatgcg tctgttcaga gatgctacgt cagcttcagc atctgtggcc
     1141 tttttctccg ccagctccag tttctcctgg gcatctttga gagcctcgga gtatttgtcc
     1201 agttcatctt cagtgccctt gagtttcttt tgcagtgaca ccagctcatc ttccagctgc
     1261 ttgctccggt cttccgcagc cttcttgtca gcctccgcct gctctgctcg atccaaggcg
     1321 ttctctttgt cgagcttcag catctgcatc ttcttcttga tggcgtccat atgtatatct
     1381 ccttcttaaa gttaaacaaa attatttcta gattcattac gaccagtcta aaaagcgcct
     1441 gaattcgcga ccttctcgtt actgacagga aaatgggcca ttggcaacca gggaaagatg
     1501 aacgtgatga tgttcacaat ttgctgaatt gtggagatct cgatcctcta cgccggacgc
     1561 atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc
     1621 accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt
     1681 atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgca tgcaccattc
     1741 cttgcggcgg cggtgctcaa cggcctcaac ctactactgg gctgcttcct aatgcaggag
     1801 tcgcataagg gagagcgtcg accgatgccc ttgagagcct tcaacccagt cagctccttc
     1861 cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa
     1921 ctcgtaggac aggtgccggc agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg
     1981 agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa
     2041 gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga agcaggccat tatcgccggc
     2101 atggcggccg acgcgctggg ctacgtcttg ctggcgttcg cgacgcgagg ctggatggcc
     2161 ttccccatta tgattcttct cgcttccggc ggcatcggga tgcccgcgtt gcaggccatg
     2221 ctgtccaggc aggtagatga cgaccatcag ggacagcttc aaggatcgct cgcggctctt
     2281 accagcctaa cttcgatcac tggaccgctg atcgtcacgg cgatttatgc cgcctcggcg
     2341 agcacatgga acgggttggc atggattgta ggcgccgccc tataccttgt ctgcctcccc
     2401 gcgttgcgtc gcggtgcatg gagccgggcc acctcgacct gaatggaagc cggcggcacc
     2461 tcgctaacgg attcaccact ccaagaattg gagccaatca attcttgcgg agaactgtga
     2521 atgcgcaaac caacccttgg cagaacatat ccatcgcgtc cgccatctcc agcagccgca
     2581 cgcggcgcat ctcgggcagc gttgggtcct ggccacgggt gcgcatgatc gtgctcctgt
     2641 cgttgaggac ccggctaggc tggcggggtt gccttactgg ttagcagaat gaatcaccga
     2701 tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga gcaacaacat
     2761 gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac gcggaagtca gcgccctgca
     2821 ccattatgtt ccggatctgc atcgcaggat gctgctggct accctgtgga acacctacat
     2881 ctgtattaac gaagcgctgg cattgaccct gagtgatttt tctctggtcc cgccgcatcc
     2941 ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg ggcatgttca tcatcagtaa
     3001 cccgtatcgt gagcatcctc tctcgtttca tcggtatcat tacccccatg aacagaaatc
     3061 ccccttacac ggaggcatca gtgaccaaac aggaaaaaac cgcccttaac atggcccgct
     3121 ttatcagaag ccagacatta acgcttctgg agaaactcaa cgagctggac gcggatgaac
     3181 aggcagacat ctgtgaatcg cttcacgacc acgctgatga gctttaccgc agctgcctcg
     3241 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     3301 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     3361 gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct
     3421 taactatgcg gcatcagagc agattgtact gagagtgcac catatatgcg gtgtgaaata
     3481 ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact
     3541 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3601 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3661 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3721 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3781 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3841 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3901 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3961 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4021 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4081 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4141 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4201 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4261 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4321 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4381 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4441 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4501 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4561 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4621 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4681 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4741 ccagttaata gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg
     4801 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4861 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4921 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4981 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5041 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac gggataatac cgcgccacat
     5101 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5161 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5221 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5281 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5341 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5401 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     5461 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     5521 cttcaagaa
//
