LOCUS       pMPS53_pME_N41_R        4153 bp    DNA     circular SYN 24-OCT-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4153)
  AUTHORS   dacer
  TITLE     Direct Submission
  JOURNAL   Exported Oct 24, 2025 from SnapGene 8.1.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4153
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      112..139
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      231..317
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     381..397
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    413..512
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(496..520)
                     /label=oAMS1186
     primer_bind     506..548
                     /label=Fragment.FOR
     primer_bind     506..523
                     /label=Fragment.FOR
     CDS             539..967
                     /codon_start=1
                     /label=N41_CNBD
                     /translation="MFYQEVRRGDFMRNWQLVAAVPLFQKLGPAVLVEIVRALRARTVP
                     AGAVICRIGESGDRMFFVVEGSVSVATPNPVELGPGAFFGEMALISGEPQSATVSAATT
                     VSLLSLHSADFQMLCSSSPEIAEIIRKTALERRGAAANA"
     misc_feature    572..574
                     /label=V12M
     misc_feature    572
                     /label=G>A
     misc_feature    704..706
                     /label=P56S
     misc_feature    704
                     /label=C>T
     primer_bind     complement(792..810)
                     /label=Primer 2
     primer_bind     811..833
                     /label=Primer 1
     primer_bind     811..833
                     /label=Primer 3
     CDS             821..823
                     /label=R307
     misc_feature    923..925
                     /label=F129I
     misc_feature    923
                     /label=T>A
     misc_feature    962..964
                     /label=S142N
     misc_feature    963
                     /label=G>A
     CDS             974..1690
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="VSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDDG
                     NYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYFG
                     FVTAAAITHGMDELYK"
     primer_bind     complement(1665..1690)
                     /label=Fragment.REV
     polyA_signal    1728..1862
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     1869..1892
                     /label=oAMS1187
     primer_bind     complement(1869..1885)
                     /label=Fragment.REV
     protein_bind    complement(1876..1975)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1993..2011)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2016..2032)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2463..3254
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      3347..3935
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
       61 caacgcaatt aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca
      121 tccgtcagga tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc
      181 cgccaccctc cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta
      241 ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga
      301 gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat
      361 gttttcccag tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc cccaaataat
      421 gattttattt tgactgatag tgacctgttc gttgcaacaa attgatgagc aatgcttttt
      481 tataatgcca actttgtaca aaaaagcagg ctccgaattc gcccttggat ccgccaccat
      541 gttctaccag gaggtgagga ggggcgactt catgaggaac tggcagctgg tggccgccgt
      601 gcctctgttc cagaagctgg gccctgccgt gctggtggag atcgtgaggg ccctgagggc
      661 caggaccgtg cctgccggcg ccgtgatctg caggatcggc gagtctggcg acaggatgtt
      721 cttcgtggtg gagggcagcg tgagcgtggc cacccctaac cctgtggagc tgggccctgg
      781 cgccttcttc ggcgagatgg ccctgatcag cggcgagcct cagagcgcca ccgtgagcgc
      841 cgccaccacc gtgagcctgc tgagcctgca cagcgccgac ttccagatgc tgtgcagcag
      901 cagccctgag atcgccgaga tcatcaggaa gaccgccctg gagaggagag gcgctgccgc
      961 taacgccgaa ttcgtgagca agggcgccga gctgttcacc ggcatcgtgc ccatcctgat
     1021 cgagctgaat ggcgatgtga atggccacaa gttcagcgtg agcggcgagg gcgagggcga
     1081 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcctgtgcc
     1141 ctggcccacc ctggtgacca ccctgagcta cggcgtgcag tgcttctcac gctaccccga
     1201 tcacatgaag cagcacgact tcttcaagag cgccatgcct gagggctaca tccaggagcg
     1261 caccatcttc ttcgaggatg acggcaacta caagtcgcgc gccgaggtga agttcgaggg
     1321 cgataccctg gtgaatcgca tcgagctgac cggcaccgat ttcaaggagg atggcaacat
     1381 cctgggcaat aagatggagt acaactacaa cgcccacaat gtgtacatca tgaccgacaa
     1441 ggccaagaat ggcatcaagg tgaacttcaa gatccgccac aacatcgagg atggcagcgt
     1501 gcagctggcc gaccactacc agcagaatac ccccatcggc gatggccctg tgctgctgcc
     1561 cgataaccac tacctgtcca cccagagcgc cctgtccaag gaccccaacg agaagcgcga
     1621 tcacatgatc tacttcggct tcgtgaccgc cgccgccatc acccacggca tggatgagct
     1681 gtacaagtga ctcgagcctc tagaactata gtgagtcgta ttacgtagat ccagacatga
     1741 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     1801 tttgtgaaat ttgtgatgct attgctttat ttggaaccat tataagctgc aataaacaag
     1861 ttaagggcga attcgaccca gctttcttgt acaaagttgg cattataaaa aataattgct
     1921 catcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc
     1981 cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct
     2041 ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgcc
     2101 tcctctagac cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt
     2161 gacgcacacc gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc
     2221 gtaagctgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac
     2281 gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg
     2341 tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga
     2401 tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca
     2461 ttatgaggga agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca
     2521 tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg
     2581 gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg
     2641 aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga
     2701 gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc
     2761 gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag
     2821 gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag
     2881 aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac
     2941 aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg
     3001 ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg
     3061 gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt
     3121 atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg
     3181 cctcgcgcgc agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg
     3241 tagtcggcaa ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt
     3301 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3361 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3421 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3481 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3541 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3601 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3661 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3721 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3781 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3841 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3901 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3961 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     4021 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     4081 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     4141 tccccgcgcg ttg
//
