LOCUS       sequence-125872-       11150 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway ORF clone of codon optimized human NF1 [NM_000267.3] with 
            stop codon (for native or N-terminal fusions).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11150)
  TITLE     Esposito lab
REFERENCE   2  (bases 1 to 11150)
  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"
COMMENT     Sequence Label: R777-E139 Hs.NF1
FEATURES             Location/Qualifiers
     source          1..11150
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..45
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     37..54
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     38..54
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    84..183
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(8641..8740)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             9117..9908
                     /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      10001..10589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10490..10509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(10919..10946)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(11038..11124)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaaa
       61 attcgagctc ggtccgagga tccgaaataa tgattttatt ttgactgata gtgacctgtt
      121 cgttgcaaca aattgataag caatgctttt ttataatgcc aactttgtac aaaaaagttg
      181 gcaccatggc cgcccacaga cctgtggaat gggtgcaggc cgtggtgtcc agattcgacg
      241 agcagctgcc catcaagacc ggacagcaga atacacacac caaggtgtcc accgagcaca
      301 acaaagagtg cctgatcaac atcagcaagt acaagttcag cctggtgatc agcggcctga
      361 ccaccatcct gaagaacgtg aacaacatgc ggatcttcgg cgaggccgcc gagaagaacc
      421 tgtacctgag ccagctgatc atcctggaca ccctggaaaa gtgcctcgct ggacagccca
      481 aggataccat gcggctggac gagacaatgc tggtgaagca gctgctgccc gagatctgcc
      541 acttcctgca cacctgccgg gagggaaatc agcacgccgc cgagctgaga aatagcgcca
      601 gcggcgtgct gttcagcctg agctgcaaca acttcaacgc cgtgttcagc cggatcagca
      661 cccggctgca ggaactgacc gtctgcagcg aggacaacgt ggacgtgcac gacatcgagc
      721 tgctgcagta catcaacgtg gactgcgcca agctgaagcg gctgctgaaa gagacagcct
      781 tcaagttcaa ggccctgaag aaggtggccc agctggccgt gatcaacagc ctggaaaaag
      841 ccttctggaa ctgggtggag aactaccccg acgagttcac caagctgtac cagatccccc
      901 agaccgacat ggccgagtgc gccgagaagc tgttcgacct ggtggacggc ttcgccgaga
      961 gcaccaagag aaaggccgcc gtgtggcccc tgcagatcat cctgctgatc ctgtgccccg
     1021 agatcatcca ggacatctcc aaggacgtgg tggacgagaa caacatgaac aagaagctgt
     1081 ttctggacag cctgaggaag gccctggccg gacacggcgg ctctagacag ctgaccgaga
     1141 gcgccgccat tgcctgcgtg aagctgtgca aggccagcac ctacatcaac tgggaggaca
     1201 acagcgtgat cttcctgctg gtgcagagca tggtggtgga cctgaagaac ctgctgttca
     1261 accccagcaa gcccttcagc agaggcagcc agcccgccga cgtggacctg atgatcgact
     1321 gcctggtgtc ctgcttccgg atcagccccc acaacaacca gcacttcaag atctgcctgg
     1381 cccagaacag ccccagcacc ttccactacg tgctggtgaa ctccctgcac cggatcatca
     1441 ccaacagcgc cctggactgg tggcccaaga tcgacgccgt gtactgccac agcgtggagc
     1501 tgcggaacat gttcggcgag acactgcaca aggccgtgca gggatgtgga gcccaccccg
     1561 ccatcagaat ggcccccagc ctgaccttca aagaaaaggt gacctccctg aagttcaaag
     1621 agaagcccac cgacctggaa acccggtcct acaagtacct gctgctgtcc atggtgaagc
     1681 tgatccacgc cgaccccaag ctgctgctgt gcaaccctag gaagcagggc cctgagacac
     1741 agggcagcac agccgagctg atcaccggcc tggtgcagct ggtgccccag agccacatgc
     1801 ctgagatcgc ccaggaagcc atggaagctc tgctcgtcct gcaccagctc gactccattg
     1861 atctgtggaa ccccgacgcc cccgtggaga cattctggga gatcagcagc cagatgctgt
     1921 tctacatctg caagaagctg accagccatc agatgctgtc cagcaccgag atcctgaagt
     1981 ggctgcggga gatcctgatc tgccggaaca agttcctgct gaagaacaag caggccgacc
     2041 ggtccagctg ccactttctg ctgttttacg gcgtgggctg cgacatcccc agcagcggca
     2101 acaccagcca gatgagcatg gaccacgagg aactgctgag aacccctggc gccagcctga
     2161 gaaagggcaa gggcaacagc agcatggaca gcgccgctgg ctgtagcggc acccccccta
     2221 tctgtcggca ggcccagaca aaactggaag tcgctctgta catgttcctg tggaatcccg
     2281 ataccgaggc cgtgctggtc gccatgagct gcttccggca cctgtgcgag gaagccgaca
     2341 tcagatgcgg cgtggatgag gtgtccgtgc acaacctgct gcccaactac aacaccttca
     2401 tggaattcgc cagcgtgagc aacatgatga gcaccggcag agccgccctg cagaaacggg
     2461 tgatggccct gctgcggaga atcgagcacc ccaccgccgg aaatacagag gcctgggagg
     2521 atacccacgc caagtgggag caggccacca agctgatcct gaactacccc aaggccaaga
     2581 tggaagatgg ccaggccgcc gaaagcctgc acaagaccat cgtgaagcgg cggatgagcc
     2641 acgtgtctgg cggcggaagc atcgacctga gcgacaccga ctccctgcaa gaatggatca
     2701 acatgaccgg ctttctgtgt gccctgggcg gagtgtgcct gcagcagcgg agcaattccg
     2761 gcctggccac ctacagccct cctatgggcc ccgtgagcga gaggaagggc agcatgatca
     2821 gcgtgatgtc cagcgagggc aacgccgaca cccccgtgag caagttcatg gaccggctgc
     2881 tgtccctgat ggtctgcaac cacgagaaag tgggcctgca gatccggacc aacgtgaagg
     2941 acctggtcgg cctggaactg agccctgccc tgtaccccat gctgttcaat aagctgaaga
     3001 ataccatctc caagttcttc gacagccagg gccaggtgct gctgaccgac accaacaccc
     3061 agttcgtgga gcagacaatc gccatcatga agaatctgct ggacaaccac accgagggca
     3121 gctctgaaca cctgggccag gccagcatcg agacaatgat gctgaacctg gtccgctacg
     3181 tgcgggtgct gggcaacatg gtgcacgcca tccagatcaa gacaaagctg tgccagctgg
     3241 tcgaggtgat gatggccaga cgggacgacc tgagcttctg ccaggaaatg aagttccgga
     3301 acaagatggt ggagtacctg accgactggg tgatgggcac cagcaaccag gccgctgacg
     3361 acgacgtgaa gtgcctgacc agggacctgg accaggcttc tatggaagct gtcgtcagtc
     3421 tgctggccgg actgcctctg cagcctgaag aaggtgacgg cgtcgagctg atggaagcca
     3481 agagccagct gttcctgaag tacttcaccc tgttcatgaa cctgctgaac gactgcagcg
     3541 aggtggagga tgagagcgcc cagaccggcg gcagaaagcg gggcatgagc agaaggctgg
     3601 cctccctgag gcactgtacc gtgctggcca tgtccaatct gctgaacgcc aacgtggata
     3661 gcggcctgat gcacagcatc ggcctgggct accacaagga cctgcagacc cgggccacct
     3721 ttatggaagt gctgaccaag atcctgcagc agggcaccga gttcgacacc ctggccgaga
     3781 ctgtgctggc cgacagattc gagagactgg tggaactggt gaccatgatg ggcgaccagg
     3841 gcgagctgcc tattgccatg gccctggcca acgtggtgcc ctgtagccag tgggatgagc
     3901 tggccagggt cctcgtcaca ctgttcgaca gccggcacct gctgtaccag ctgctgtgga
     3961 atatgttcag caaagaggtg gagctggccg actccatgca gaccctgttc agaggcaact
     4021 ccctggccag caagatcatg accttttgct tcaaggtgta cggcgccacc tacctgcaga
     4081 agctgctgga ccccctgctg aggatcgtga tcaccagcag cgactggcag cacgtgagct
     4141 tcgaggtgga ccccaccaga ctggaaccca gcgagagcct ggaagagaac cagcggaacc
     4201 tgctgcagat gaccgagaag ttcttccacg ccatcatcag cagcagctcc gagttccccc
     4261 cccagctgag aagcgtgtgc cactgcctgt accaggtggt gtcccagcgg ttcccccaga
     4321 atagcatcgg cgccgtgggc agcgccatgt tcctgcggtt catcaaccct gccatcgtga
     4381 gcccttacga ggccggcatc ctggataaga agccccctcc ccggatcgag aggggcctga
     4441 agctgatgag caagattctg cagagcattg ccaaccacgt gctgtttacc aaggaagagc
     4501 acatgcggcc cttcaacgac ttcgtgaaga gcaacttcga cgccgccagg cggttcttcc
     4561 tggatatcgc cagcgactgc cctaccagcg acgccgtgaa ccacagcctg tccttcatca
     4621 gcgacggcaa cgtcctggcc ctgcatagac tgctgtggaa caaccaggaa aagatcggcc
     4681 agtacctgag cagcaaccgg gaccacaagg ctgtgggcag gcggcccttc gataagatgg
     4741 ccaccctgct ggcctatctg ggccctcccg agcacaagcc tgtggccgac acccattggt
     4801 ctagcctgaa cctgaccagc agcaagttcg aggaattcat gacccggcac caggtgcacg
     4861 agaaagaaga gtttaaagcc ctgaaaaccc tgagcatctt ctaccaggcc ggcacctcca
     4921 aggccggcaa ccccatcttc tactacgtgg ccagaaggtt caagacaggc cagatcaacg
     4981 gcgacctgct gatctaccac gtgctgctga cactgaagcc ctactacgcc aagccctacg
     5041 agatcgtggt ggatctgacc cacaccggcc ccagcaaccg gttcaagacc gacttcctga
     5101 gcaagtggtt cgtggtgttc cccggcttcg cctacgataa cgtgagcgcc gtgtacatct
     5161 acaactgcaa cagctgggtg cgggagtaca ccaagtacca cgagcgcctg ctgaccggcc
     5221 tgaagggctc caagcggctg gtgttcatcg actgccccgg caagctggcc gagcacatcg
     5281 agcacgagca gcagaaactg cctgccgcca ccctggccct ggaagaggac ctgaaggtgt
     5341 tccacaacgc cctgaagctg gcccacaagg atacaaaggt gtccatcaaa gtgggcagca
     5401 ccgccgtgca ggtgaccagc gccgagagga caaaggtgct gggccagagc gtgttcctga
     5461 acgacatcta ctacgcctcc gagatcgagg aaatctgcct ggtggatgag aaccagttca
     5521 ccctgactat cgccaaccag ggcacccctc tgaccttcat gcaccaggaa tgcgaggcca
     5581 tcgtccagtc catcatccac atccggacca gatgggagct gtcccagccc gacagcatcc
     5641 cccagcacac caagatccgg cccaaggatg tgcccggcac cctgctgaat atcgctctgc
     5701 tgaatctggg cagcagcgac ccttctctgc ggagcgccgc ctacaatctg ctgtgcgccc
     5761 tgacctgcac cttcaacctg aagatcgagg gccagctgct ggaaaccagc ggcctgtgca
     5821 tccccgccaa caacaccctg tttatcgtga gcatcagcaa gaccctggcc gccaacgagc
     5881 ctcacctgac cctggaattt ctggaagagt gcatcagcgg cttcagcaag agcagcatcg
     5941 agctgaagca cctgtgcctg gaatacatga ccccctggct gtctaatctg gtccggttct
     6001 gcaagcacaa cgacgacgcc aagcggcaga gagtgaccgc cattctggac aagctgatca
     6061 caatgaccat caacgagaag cagatgtacc ccagcatcca ggccaagatc tggggcagcc
     6121 tgggccagat caccgatctg ctggatgtgg tgctggacag cttcatcaag accagcgcca
     6181 ccggcggact gggaagcatc aaggccgagg tgatggccga tacagccgtg gccctggcta
     6241 gcggcaacgt gaagctggtg tccagcaagg tgatcggccg gatgtgcaag atcatcgaca
     6301 agacctgcct gagccccacc cctaccctgg aacagcacct gatgtgggac gatatcgcca
     6361 tcctggcccg gtacatgctg atgctgtcct tcaacaatag cctggacgtg gccgcccatc
     6421 tgccctacct gttccacgtg gtgacctttc tggtggccac cggccctctg agcctgagag
     6481 ccagcaccca cggcctggtg attaacatca tccactccct gtgcacctgc tcccagctgc
     6541 acttcagcga ggaaaccaag caggtgctga gactgagcct gaccgagttc tccctgccca
     6601 agttctacct gctgtttggc atcagcaaag tgaagtctgc cgccgtgatc gccttcaggt
     6661 ccagctacag ggacagatct ttctcccccg gaagctacga gagagagaca ttcgccctga
     6721 caagcctgga aaccgtgaca gaggccctgc tggaaatcat ggaagcctgc atgagagaca
     6781 tccccacctg caagtggctc gaccagtgga ccgaactggc ccagaggttc gccttccagt
     6841 acaaccccag cctgcagcct agagccctgg tggtgttcgg ctgcatctcc aagagagtga
     6901 gccacggcca gattaagcag atcatcagga tcctgtccaa ggccctggaa agctgtctga
     6961 agggccccga cacctacaac tcccaggtgc tgatcgaggc cacagtgatc gccctgacca
     7021 aactgcagcc actgctgaat aaggacagcc ccctgcacaa agccctgttc tgggtggccg
     7081 tggccgtgct gcagctggac gaggtgaacc tgtacagcgc cgggaccgct ctgctggaac
     7141 agaatctgca caccctggac tccctgagaa tcttcaacga caagagcccc gaggaagtgt
     7201 tcatggccat ccggaacccc ctggaatggc actgcaagca gatggaccac ttcgtgggcc
     7261 tgaacttcaa cagcaacttc aacttcgccc tggtcggaca cctgctgaag ggctacagac
     7321 accccagccc agccatcgtg gccaggaccg tgcggatcct gcataccctg ctgaccctgg
     7381 tgaacaagca ccggaactgc gacaagtttg aggtgaacac ccagagcgtg gcctacctgg
     7441 ctgccctgct gacagtgagc gaggaagtgc ggagccggtg cagcctgaag cacaggaaga
     7501 gcctgctcct cactgatatc agcatggaaa acgtgcccat ggacacctac cccatccacc
     7561 acggcgaccc cagctacagg accctgaaag aaacccagcc ctggtcctcc cctaagggca
     7621 gcgagggcta tctggccgcc acctatccta ccgtcggcca gacaagccct agggcccgca
     7681 agtccatgag cctggacatg ggccagccta gccaggccaa caccaagaaa ctgctgggca
     7741 cccggaagag cttcgaccac ctgatcagcg acaccaaggc ccccaagagg caggaaatgg
     7801 aaagcggcat caccaccccc cccaagatgc ggagagtggc cgagacagac tacgagatgg
     7861 aaacccagcg gatcagcagc tcccagcagc accctcacct gcggaaagtg tccgtgtccg
     7921 agagcaatgt gctgctggac gaagaggtgc tgaccgatcc taagatccag gccctgctgc
     7981 tgactgtcct ggccaccctg gtgaagtaca ccaccgacga gttcgatcag cggatcctgt
     8041 acgagtacct ggccgaggcc agcgtggtgt ttcccaaggt gttccctgtg gtgcacaatc
     8101 tgctggatag caagatcaac accctgctgt ctctgtgcca ggaccccaac ctgctgaatc
     8161 ccatccacgg catcgtgcag agtgtggtgt accacgaaga gagcccccct cagtaccaga
     8221 ccagctacct gcagtccttc ggcttcaacg gcctgtggcg gtttgccggc cctttcagca
     8281 agcagaccca gatccccgac tacgccgaac tgatcgtgaa gttcctggac gccctgatcg
     8341 atacctacct gcccggcatc gacgaggaaa cctccgagga atctctgctg acccccacct
     8401 ccccttatcc tccagccctg cagtcccagc tgtccatcac cgccaacctg aacctgtcca
     8461 actccatgac cagcctggcc acatctcagc acagccctgg catcgacaaa gaaaacgtgg
     8521 agctgtctcc caccaccggc cactgcaact ccggcaggac cagacacgga agcgccagcc
     8581 aggtgcagaa gcagagaagc gccggcagct tcaagcggaa cagcatcaag aaaatcgtgt
     8641 aatcaacttt cttgtacaaa gttggcatta taggaaagca ttgcttatca atttgttgca
     8701 acgaacaggt cactatcagt caaaataaaa tcattatttg catgcaagct tagatcctct
     8761 agaccagcca ggacagaaat gcctcgactt cgctgctacc caaggttgcc gggtgacgca
     8821 caccgtggaa acggatgaag gcacgaaccc agtggacata agcctgttcg gttcgtaagc
     8881 tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg
     8941 gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt ggggtacagt
     9001 ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat
     9061 ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta aacattatga
     9121 gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc gtcatcgagc
     9181 gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg gatggcggcc
     9241 tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa
     9301 cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga gagagcgaga
     9361 ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg tggcgttatc
     9421 cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt gcaggtatct
     9481 tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca agagaacata
     9541 gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct gaacaggatc
     9601 tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac tgggctggcg
     9661 atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta accggcaaaa
     9721 tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc cagtatcagc
     9781 ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc ttggcctcgc
     9841 gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc aaggtagtcg
     9901 gcaaataacc ctcgagccac ccatgaccaa aatcccttaa cgtgagttac gcgtcgttcc
     9961 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
    10021 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
    10081 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
    10141 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
    10201 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
    10261 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
    10321 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
    10381 tacagcgtga gcattgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
    10441 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
    10501 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
    10561 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
    10621 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
    10681 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
    10741 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
    10801 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
    10861 gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta gaaacgcaaa
    10921 aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt atggcgggcg
    10981 tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc ccggcggatt
    11041 tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttc
    11101 cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg
//
