LOCUS       sequence-486891-        7711 bp    DNA     circular SYN 23-SEP-2025
DEFINITION  Bacterial Expression of HsBGG-WT.
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 Sep 23, 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-WT_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"
     primer_bind     669..685
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     670..686
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(702..718)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(702..718)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(751..1108)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      765..900
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        917..1108
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     1065..1085
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    2678..3072
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     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 tgggtaccgg
      661 gccccccctc gaggtcgacg gtatcgataa gcttgatggg ggatccacta gttctagagc
      721 ggccgcactc tagcctcgag gctagagctt tttgcaaaag cctaggcctc caaaaaagcc
      781 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
      841 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
      901 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact
      961 tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat
     1021 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac
     1081 attccacagc tggttctttc cgcctcagaa ggtaccggat cttgccattg cactccagcc
     1141 tggacaacaa gagcaaaact ccatctaaaa aaaaaaaaaa atctactctc tttgcaaatt
     1201 ttaaatatac ttacagtaca attaactaca gtcacgatgc tgtacattag atatctagaa
     1261 tttattcatc ccacataact gaaattttat accctttgag taatagtttc ctgattctcc
     1321 cacccccaac ccctggaaac catacctcta ctgtctgttt ccatgagagt gactttttta
     1381 tgtttcacat atgagcaagg ttgtgtagta tttgtctttt tgtgtcaggc ttatttttct
     1441 tagttttaca tcctccaggt tcatcaattc tgccataaat ggcaggattt tcttcttttt
     1501 aattttttta ttttcctatt tttaattttt tgtttgttat tgaatcatct gtgtttctta
     1561 tatattttgg atattaacca aaatatttaa gaaatatttg gtttgcaaat atgccatttc
     1621 atggttcacc tttcatttgt tcattgtttt gttttttgtg tggaaatttt ttagtttgat
     1681 gtagcctcac ttctttattt ttccttttgt tgcctttgct tctcatatca aattcaaaaa
     1741 acggctgcaa catgaatatt agaattttct tcttttgatt tctttaaaaa tttttatggt
     1801 tttaggtctt agtttaattc tttaacccat tcaaatttat ttttgtttat ggtgtaagac
     1861 aagggtctga tttcatttca ttttgagggt ttcaccctct catataataa aattcttcta
     1921 tatacccttc tatcatcact tattactgat tatatgcctt gatttcagaa tcttgccaac
     1981 tagattcaga atttgactgg gagagaggac aaggaccact tgagactcat attttatttc
     2041 cagaatctag catctaccta cctaggtatt gaataagaaa aatgaagtta aggtggttga
     2101 tggtaacact atgctaataa ctgcagagcc agaagcacca taagggacat gataagggag
     2161 ccagcagacc tctgatctct tcctgaatgc taatcttaaa catcctgagg aagaatggga
     2221 cttccatttg gggtgggcct atgacagggt aataagacag tagtgaatat caagctacaa
     2281 aaagccccct ttcaaattct tctcagtcct aacttttcat actaagccca gtccttccaa
     2341 agcagactgt gaaagagtga tagttccggg agactagcac tgcagattcc gggtcactgt
     2401 gagtggggga ggcagggaag aagggctcac aggacagtca aaccatgacc cctgtttttc
     2461 cttcttcaag tagacctcta taagacaaca gagacaacta aggctgagtg gccaggcgag
     2521 gagaaaccat ctcgccgtaa aacatggaag gaacacttca ggggaaaggt ggtatctcta
     2581 agcaagagaa ctgagtggag acaaggctga gagatgcagg ataagcaaat gggtagtgaa
     2641 aagacattca tgaggacagc taaaacaata agtaatgtaa aatacagcat agcaaaactt
     2701 taacctccaa atcaagcctc tacttgaatc cttttctgag ggatgaataa ggcataggca
     2761 tcaggggctg ttgccaatgt gcattagctg tttgcagcct caccttcttt catggagttt
     2821 aagatatagt gtattttccc aaggtttgaa ctagctcttc atttctttat gttttaaatg
     2881 cactgacctc ccacattccc tttttagtaa aatattcaga aataatttaa atacatcatt
     2941 gcaatgaaaa taaatgtttt ttattaggca gaatccagat gctcaaggcc cttcataata
     3001 tcccccagtt tagtagttgg acttagggaa caaaggaacc tttaatagaa attggacagc
     3061 aagaaagcga gcttagtgat acttgtgggc cagggcatta gccacaccag ccaccacttt
     3121 ctgataggca gcctgcactg gtggggtgaa ttctttgcca aagtgatggg ccagcacaca
     3181 gaccagcacg ttgcccagga gctgtgggag gaagataaga ggtatgaaca tgattagcaa
     3241 aagggcctag cttggactca gaataatcca gccttatccc aaccataaaa taaaagcaga
     3301 atggtagctg gattgtagct gctattagca atatgaaacc tcttacatca gttacaattt
     3361 atatgcagaa atatttatat gcagaaatat tgctattgcc ttaacccaga aattatcact
     3421 gttattcttt agaatggtgc aaagaggcat gatacattgt atcattattg ccctgaaaga
     3481 aagagattag ggaaagtatt agaaataaga taaacaaaaa agtatattaa aagaagaaag
     3541 cattttttaa aattacaaat gcaaaattac cctgatttgg tcaatatgtg tacacatatt
     3601 aaaacattac actttaaccc ataaatatgt ataatgatta tgtatcaatt aaaaataaaa
     3661 gaaaataaag tagggagatt atgaatatgc aaataagcac acatatattc caaatagtaa
     3721 tgtactaggc agactgtgta aagttttttt ttaagttact taatgtatct cagagatatt
     3781 tccttttgtt atacacaatg ttaaggcatt aagtataata gtaaaaattg cggagaagaa
     3841 aaaaaaagaa agcaagaatt aaacaaaaga aaacaattgt tatgaacagc aaataaaaga
     3901 aactaaaacg atcctgagac ttccacactg atgcaatcat tcgtctgttt cccattctaa
     3961 actgtaccct gttacttatc cccttcctat gacatgaact taaccataga aaagaagggg
     4021 aaagaaaaca tcaagggtcc catagactca ccctgaagtt ctcaggatcc acgtgcagct
     4081 tgtcacagtg cagctcactc agtgtggcaa aggtgccctt gaggttgtcc aggtgagcca
     4141 ggccatcact aaaggcaccg agcactttct tgccatgagc cttcacctta gggttgccca
     4201 taacagcatc aggagtggac agatccccaa aggactcaaa gaacctctgg gtccaagggt
     4261 agaccaccag cagcctaagg gtgggaaaat agaccaatag gcagagagag tcagtgccta
     4321 tcagaaaccc aagagtcttc tctgtctcca catgcccagt ttctattggt ctccttaaac
     4381 ctgtcttgta accttgatac caacctgccc agggcctcac caccaacttc atccacgttc
     4441 accttgcccc acagggcagt aacggcagac ttctcctcag gagtcaggtg caccatggtg
     4501 tctgtttgag gttgctagtg aacacagttg tgtcagaagc aaatgtaagc aatagatggc
     4561 tctgccctga cttttatgcc cagccctggc tcctgccctc cctgctcctg ggagtagatt
     4621 ggccaaccct agggtgtggc tccacagggt gaggtctaag tgatgacagc cgtacctgtc
     4681 cttggctctt ctggcactgg cttaggagtt ggacttcaaa ccctcagccc tccctctaag
     4741 atatatctct tggccccata ccatcagtac aaattgctac taaaaacatc ctcctttgca
     4801 agtgtattta cgtaatattt ggaatcacag cttggtaagc atattgaaga tcgttttccc
     4861 aattttctta ttacacaaat aagaaattga tgcactaaaa gtggaagagt tttgtctacc
     4921 ataattcagc tttgggatat gtagatggat ctcttcctgc gtctccagaa tatgcaaaat
     4981 acttacagga cagaatggat gaaaactcta cctcagttct aagcatatct tctccttatt
     5041 tggattaaaa ccttctggta agaaaagaaa aaaaatatat atatatatgt gtatatatac
     5101 acacatacat atacatatat atgcattcat ttgttgttgt ttttcttaat ttgctcatgc
     5161 atgctaataa attatgtcta aaaatagaat aaatacaaat caatgtgctc tgtgcattag
     5221 ttacttatta ggttttggga aacaagagat aaaaaactag agacctctta atgcagtcaa
     5281 aaatacaaat aaataaaaag tcacttacaa cccaaagtgt gactatcaat ggggtaatca
     5341 gtggtgtcaa ataggaggtt aaccaaaaga aactggatcc tctatttcta gttatcagaa
     5401 ggaaatttac aaatttctta tttccattgc tttattctct taaatgcttt ctctattatt
     5461 gctaaataaa tagagatccc cgatccgcgg ccgccaccgc ggtggagctc 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 gaagaacagt 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
//
