LOCUS       sequence-288631-        7711 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of HsBGG-PTC.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7711)
  AUTHORS   Gatfield D, Unterholzner L, Ciccarelli FD, Bork P, Izaurralde E
  TITLE     Nonsense-mediated mRNA decay in Drosophila: at the intersection of 
            the yeast and mammalian pathways.
  JOURNAL   EMBO J. 2003 Aug 1. 22(15):3960-70.
  PUBMED    12881430
REFERENCE   2  (bases 1 to 7711)
  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: "EMBO J. 
            2003 Aug 1. 22(15):3960-70."
COMMENT     Sequence Label: pBSII(KSplus)-HsBGG-PTC_E
FEATURES             Location/Qualifiers
     source          1..7711
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3091..3485
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     promoter        5055..5412
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5055..5246
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5078..5098)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5263..5398
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5445..5461
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5445..5461
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5477..5493)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5478..5494)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(5523..5543)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5523..5541)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5562..5578)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5562..5578)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5575..5597)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5586..5602
                     /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(5610..5616,5617..5634,5635..5640))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5655..5676
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5793..5810)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5964..6552)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6044..6063)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6723..7514,7515..7583))
                     /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     7346..7365
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7584..7688)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctcca
      661 ccgcggtggc ggccgcggat cggggatctc tatttattta gcaataatag agaaagcatt
      721 taagagaata aagcaatgga aataagaaat ttgtaaattt ccttctgata actagaaata
      781 gaggatccag tttcttttgg ttaacctcct atttgacacc actgattacc ccattgatag
      841 tcacactttg ggttgtaagt gactttttat ttatttgtat ttttgactgc attaagaggt
      901 ctctagtttt ttatctcttg tttcccaaaa cctaataagt aactaatgca cagagcacat
      961 tgatttgtat ttattctatt tttagacata atttattagc atgcatgagc aaattaagaa
     1021 aaacaacaac aaatgaatgc atatatatgt atatgtatgt gtgtatatat acacatatat
     1081 atatatattt tttttctttt cttaccagaa ggttttaatc caaataagga gaagatatgc
     1141 ttagaactga ggtagagttt tcatccattc tgtcctgtaa gtattttgca tattctggag
     1201 acgcaggaag agatccatct acatatccca aagctgaatt atggtagaca aaactcttcc
     1261 acttttagtg catcaatttc ttatttgtgt aataagaaaa ttgggaaaac gatcttcaat
     1321 atgcttacca agctgtgatt ccaaatatta cgtaaataca cttgcaaagg aggatgtttt
     1381 tagtagcaat ttgtactgat ggtatggggc caagagatat atcttagagg gagggctgag
     1441 ggtttgaagt ccaactccta agccagtgcc agaagagcca aggacaggta cggctgtcat
     1501 cacttagacc tcaccctgtg gagccacacc ctagggttgg ccaatctact cccaggagca
     1561 gggagggcag gagccagggc tgggcataaa agtcagggca gagccatcta ttgcttacat
     1621 ttgcttctga cacaactgtg ttcactagca acctcaaaca gacaccatgg tgcacctgac
     1681 tcctgaggag aagtctgccg ttactgccct gtggggcaag gtgaacgtgg atgaagttgg
     1741 tggtgaggcc ctgggcaggt tggtatcaag gttacaagac aggtttaagg agaccaatag
     1801 aaactgggca tgtggagaca gagaagactc ttgggtttct gataggcact gactctctct
     1861 gcctattggt ctattttccc acccttaggc tgctggtggt ctacccttgg acctagagat
     1921 tctttgagtc ctttggggat ctgtccactc ctgatgctgt tatgggcaac cctaaggtga
     1981 aggctcatgg caagaaagtg ctcggtgcct ttagtgatgg cctggctcac ctggacaacc
     2041 tcaagggcac ctttgccgca ctgagtgagc tgcactgtga caagctgcac gtggatcctg
     2101 agaacttcag ggtgagtcta tgggaccctt gatgttttct ttccccttct tttctatggt
     2161 taagttcatg tcataggaag gggagaagta acagggtaca gtttagaatg ggaaacagac
     2221 gaatgattgc atcagtgtgg aagtctcagg atcgttttag tttcttttat ttgctgttca
     2281 taacaattgt tttcttttgt ttaattcttg ctttcttttt ttttcttctc cgcaattttt
     2341 actattatac ttaatgcctt aacattgtgt ataacaaaag gaaatatctc tgagatacat
     2401 taagtaactt aaaaaaaaac tttacacagt ctgcctagta cattactatt tggaatatat
     2461 gtgtgcttat ttgcatattc ataatctccc tactttattt tcttttattt ttaattgata
     2521 cataatcatt atacatattt atgggttaaa gtgtaatgtt ttaatatgtg tacacatatt
     2581 gaccaaatca gggtaatttt gcatttgtaa ttttaaaaaa tgctttcttc ttttaatata
     2641 cttttttgtt tatcttattt ctaatacttt ccctaatctc tttctttcag ggcaataatg
     2701 atacaatgta tcatgcctct ttgcaccatt ctaaagaata acagtgataa tttctgggtt
     2761 aaggcaatag caatatttct gcatataaat atttctgcat ataaattgta actgatgtaa
     2821 gaggtttcat attgctaata gcagctacaa tccagctacc attctgcttt tattttatgg
     2881 ttgggataag gctggattat tctgagtcca agctaggccc ttttgctaat catgttcata
     2941 cctcttatct tcctcccaca gctcctgggc aacgtgctgg tctgtgtgct ggcccatcac
     3001 tttggcaaag aattcacccc accagtgcag gctgcctatc agaaagtggt ggctggtgtg
     3061 gctaatgccc tggcccacaa gtatcactaa gctcgctttc ttgctgtcca atttctatta
     3121 aaggttcctt tgttccctaa gtccaactac taaactgggg gatattatga agggccttga
     3181 gcatctggat tctgcctaat aaaaaacatt tattttcatt gcaatgatgt atttaaatta
     3241 tttctgaata ttttactaaa aagggaatgt gggaggtcag tgcatttaaa acataaagaa
     3301 atgaagagct agttcaaacc ttgggaaaat acactatatc ttaaactcca tgaaagaagg
     3361 tgaggctgca aacagctaat gcacattggc aacagcccct gatgcctatg ccttattcat
     3421 ccctcagaaa aggattcaag tagaggcttg atttggaggt taaagttttg ctatgctgta
     3481 ttttacatta cttattgttt tagctgtcct catgaatgtc ttttcactac ccatttgctt
     3541 atcctgcatc tctcagcctt gtctccactc agttctcttg cttagagata ccacctttcc
     3601 cctgaagtgt tccttccatg ttttacggcg agatggtttc tcctcgcctg gccactcagc
     3661 cttagttgtc tctgttgtct tatagaggtc tacttgaaga aggaaaaaca ggggtcatgg
     3721 tttgactgtc ctgtgagccc ttcttccctg cctcccccac tcacagtgac ccggaatctg
     3781 cagtgctagt ctcccggaac tatcactctt tcacagtctg ctttggaagg actgggctta
     3841 gtatgaaaag ttaggactga gaagaatttg aaagggggct ttttgtagct tgatattcac
     3901 tactgtctta ttaccctgtc ataggcccac cccaaatgga agtcccattc ttcctcagga
     3961 tgtttaagat tagcattcag gaagagatca gaggtctgct ggctccctta tcatgtccct
     4021 tatggtgctt ctggctctgc agttattagc atagtgttac catcaaccac cttaacttca
     4081 tttttcttat tcaataccta ggtaggtaga tgctagattc tggaaataaa atatgagtct
     4141 caagtggtcc ttgtcctctc tcccagtcaa attctgaatc tagttggcaa gattctgaaa
     4201 tcaaggcata taatcagtaa taagtgatga tagaagggta tatagaagaa ttttattata
     4261 tgagagggtg aaaccctcaa aatgaaatga aatcagaccc ttgtcttaca ccataaacaa
     4321 aaataaattt gaatgggtta aagaattaaa ctaagaccta aaaccataaa aatttttaaa
     4381 gaaatcaaaa gaagaaaatt ctaatattca tgttgcagcc gttttttgaa tttgatatga
     4441 gaagcaaagg caacaaaagg aaaaataaag aagtgaggct acatcaaact aaaaaatttc
     4501 cacacaaaaa agaaaacaat gaacaaatga aaggtgaacc atgaaatggc atatttgcaa
     4561 accaaatatt tcttaaatat tttggttaat atccaaaata tataagaaac acagatgatt
     4621 caataacaaa caaaaaatta aaaataggaa aataaaaaaa ttaaaaagaa gaaaatcctg
     4681 ccatttatgg cagaattgat gaacctggag gatgtaaaac taagaaaaat aagcctgaca
     4741 caaaaagaca aatactacac aaccttgctc atatgtgaaa cataaaaaag tcactctcat
     4801 ggaaacagac agtagaggta tggtttccag gggttggggg tgggagaatc aggaaactat
     4861 tactcaaagg gtataaaatt tcagttatgt gggatgaata aattctagat atctaatgta
     4921 cagcatcgtg actgtagtta attgtactgt aagtatattt aaaatttgca aagagagtag
     4981 atttttttgt ttttttagat ggagttttgc tcttgttgtc caggctggag tgcaatggca
     5041 agatccggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     5101 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5161 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5221 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     5281 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5341 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     5401 ggcttttgca aaaagctcta gcctcgaggc tagagtgcgg ccgctctaga actagtggat
     5461 cccccatcaa gcttatcgat accgtcgacc tcgagggggg gcccggtacc cagcttttgt
     5521 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
     5581 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     5641 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     5701 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     5761 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5821 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5881 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5941 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6001 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6061 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6121 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6181 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6241 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6301 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6361 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     6421 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6481 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6541 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6601 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6661 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6721 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6781 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6841 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6901 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6961 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7021 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7081 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7141 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7201 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7261 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7321 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7381 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7441 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7501 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7561 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7621 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7681 gttccgcgca catttccccg aaaagtgcca c
//
