LOCUS       pMPS15_pME_N41_a        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      54..81
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      173..259
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     323..339
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    355..454
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(438..462)
                     /label=oAMS1186
     primer_bind     448..490
                     /label=Fragment.FOR
     primer_bind     448..465
                     /label=Fragment.FOR
     CDS             481..909
                     /codon_start=1
                     /label=N41_CNBD
                     /translation="MFYQEVRRGDFMRNWQLVAAVPLFQKLGPAVLVEIVRALRARTVP
                     AGAVICRIGESGDRMFFVVEGSVSVATPNPVELGPGAFFGEMALISGEPRSATVSAATT
                     VSLLSLHSADFQMLCSSSPEIAEIIRKTALERRGAAANA"
     misc_feature    514..516
                     /label=V12M
     misc_feature    514
                     /label=G>A
     misc_feature    646..648
                     /label=P56S
     misc_feature    646
                     /label=C>T
     primer_bind     complement(734..752)
                     /label=Primer 2
     primer_bind     753..775
                     /label=Primer 1
     primer_bind     753..775
                     /label=Primer 3
     CDS             763..765
                     /label=R307
     misc_feature    865..867
                     /label=F129I
     misc_feature    865
                     /label=T>A
     misc_feature    904..906
                     /label=S142N
     misc_feature    905
                     /label=G>A
     CDS             916..1632
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="VSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDDG
                     NYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYFG
                     FVTAAAITHGMDELYK"
     primer_bind     complement(1607..1632)
                     /label=Fragment.REV
     polyA_signal    1670..1804
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     1811..1834
                     /label=oAMS1187
     primer_bind     complement(1811..1827)
                     /label=Fragment.REV
     protein_bind    complement(1818..1917)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1935..1953)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1958..1974)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2405..3196
                     /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      3289..3877
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cgcaacgcaa ttaatacgcg taccgctagc caggaagagt ttgtagaaac gcaaaaaggc
       61 catccgtcag gatggccttc tgcttagttt gatgcctggc agtttatggc gggcgtcctg
      121 cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc
      181 tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag tcttccgact
      241 gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcgttaac gctagcatgg
      301 atgttttccc agtcacgacg ttgtaaaacg acggccagtc ttaagctcgg gccccaaata
      361 atgattttat tttgactgat agtgacctgt tcgttgcaac aaattgatga gcaatgcttt
      421 tttataatgc caactttgta caaaaaagca ggctccgaat tcgcccttgg atccgccacc
      481 atgttctacc aggaggtgag gaggggcgac ttcatgagga actggcagct ggtggccgcc
      541 gtgcctctgt tccagaagct gggccctgcc gtgctggtgg agatcgtgag ggccctgagg
      601 gccaggaccg tgcctgccgg cgccgtgatc tgcaggatcg gcgagtctgg cgacaggatg
      661 ttcttcgtgg tggagggcag cgtgagcgtg gccaccccta accctgtgga gctgggccct
      721 ggcgccttct tcggcgagat ggccctgatc agcggcgagc ctaggagcgc caccgtgagc
      781 gccgccacca ccgtgagcct gctgagcctg cacagcgccg acttccagat gctgtgcagc
      841 agcagccctg agatcgccga gatcatcagg aagaccgccc tggagaggag aggcgctgcc
      901 gctaacgccg aattcgtgag caagggcgcc gagctgttca ccggcatcgt gcccatcctg
      961 atcgagctga atggcgatgt gaatggccac aagttcagcg tgagcggcga gggcgagggc
     1021 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcctgtg
     1081 ccctggccca ccctggtgac caccctgagc tacggcgtgc agtgcttctc acgctacccc
     1141 gatcacatga agcagcacga cttcttcaag agcgccatgc ctgagggcta catccaggag
     1201 cgcaccatct tcttcgagga tgacggcaac tacaagtcgc gcgccgaggt gaagttcgag
     1261 ggcgataccc tggtgaatcg catcgagctg accggcaccg atttcaagga ggatggcaac
     1321 atcctgggca ataagatgga gtacaactac aacgcccaca atgtgtacat catgaccgac
     1381 aaggccaaga atggcatcaa ggtgaacttc aagatccgcc acaacatcga ggatggcagc
     1441 gtgcagctgg ccgaccacta ccagcagaat acccccatcg gcgatggccc tgtgctgctg
     1501 cccgataacc actacctgtc cacccagagc gccctgtcca aggaccccaa cgagaagcgc
     1561 gatcacatga tctacttcgg cttcgtgacc gccgccgcca tcacccacgg catggatgag
     1621 ctgtacaagt gactcgagcc tctagaacta tagtgagtcg tattacgtag atccagacat
     1681 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     1741 tatttgtgaa atttgtgatg ctattgcttt atttggaacc attataagct gcaataaaca
     1801 agttaagggc gaattcgacc cagctttctt gtacaaagtt ggcattataa aaaataattg
     1861 ctcatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     1921 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     1981 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     2041 cctcctctag accagccagg acagaaatgc ctcgacttcg ctgctaccca aggttgccgg
     2101 gtgacgcaca ccgtggaaac ggatgaaggc acgaacccag tggacataag cctgttcggt
     2161 tcgtaagctg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga
     2221 acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgg
     2281 ggtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt
     2341 gatgttatgg agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttaaa
     2401 cattatgagg gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt
     2461 catcgagcgc catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga
     2521 tggcggcctg aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga
     2581 tgaaacaacg cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga
     2641 gagcgagatt ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg
     2701 gcgttatcca gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc
     2761 aggtatcttc gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag
     2821 agaacatagc gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga
     2881 acaggatcta tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg
     2941 ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac
     3001 cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca
     3061 gtatcagccc gtcatacttg aagctagaca ggcttatctt ggacaagaag aagatcgctt
     3121 ggcctcgcgc gcagatcagt tggaagaatt tgtccactac gtgaaaggcg agatcaccaa
     3181 ggtagtcggc aaataaccct cgagccaccc atgaccaaaa tcccttaacg tgagttacgc
     3241 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3301 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3361 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3421 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3481 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3541 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3601 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3661 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3721 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3781 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     3841 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3901 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     3961 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     4021 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     4081 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     4141 agcgggcagt gag
//
