LOCUS       sequence-229901-       10788 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transfer vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10788)
  AUTHORS   Cui Y, Morrison H
  TITLE     Construction of cloning-friendly minigenes for mammalian expression 
            of full-length human NF1 isoforms.
  JOURNAL   Hum Mutat. 2018 Nov 8. doi: 10.1002/humu.23681.
  PUBMED    30408279
REFERENCE   2  (bases 1 to 10788)
  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: "Hum Mutat. 
            2018 Nov 8. doi: 10.1002/humu.23681."
COMMENT     Sequence Label: pAMP-CY1_NF1 T2 (+minintr, KDR)
FEATURES             Location/Qualifiers
     source          1..10788
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     103..122
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(9013..9035)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9180..9198)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9266..9370
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9371..9439,9440..10231)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(9589..9608)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(10402..10788,1..202)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
       61 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
      121 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
      181 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
      241 ttgctggcct tttgctcaca tgtcgtacga cgcgtcccgg gaccggtggc atggccgcgc
      301 acaggccggt ggaatgggtc caggccgtgg tcagccgctt cgacgagcag cttccaataa
      361 aaacaggaca gcagaacaca cataccaaag tcagtactga gcacaacaag gaatgtctaa
      421 tcaatatttc caaatacaag ttttctttgg ttataagcgg cctcactact attttaaaga
      481 atgttaacaa tatgagaata tttggagaag ctgctgaaaa aaatttatat ctctctcagt
      541 tgattatatt ggatacactg gaaaaatgtc ttgctgggca accaaaggac acaatgagat
      601 tagatgaaac gatgctggtc aaacagttgc tgccagaaat ctgccatttt cttcacacct
      661 gtcgtgaagg aaaccagcat gcagctgaac ttcggaattc tgcctctggg gttttatttt
      721 ctctcagctg caacaacttc aatgcagtct ttagtcgcat ttctaccagg ttacaggaat
      781 taactgtttg ttcagaagac aatgttgatg ttcatgatat agaattgtta cagtatatca
      841 atgtggattg tgcaaaatta aaacgactcc tgaaggaaac agcatttaaa tttaaagccc
      901 taaagaaggt tgcgcagtta gcagttataa atagcctgga aaaggcattt tggaactggg
      961 tagaaaatta tccagatgaa tttacaaaac tataccagat cccacagact gatatggctg
     1021 aatgtgcaga aaagctattt gacttggtgg atggttttgc tgaaagcacc aaacgtaaag
     1081 cagcagtttg gccactacaa atcattctcc ttatcttgtg tccagaaata atccaggata
     1141 tatccaaaga cgtggttgat gaaaacaaca tgaataagaa gttatttctg gacagtctac
     1201 gaaaagctct tgctggccat ggaggaagta ggcagctgac agaaagtgct gcaattgcct
     1261 gtgtcaaact gtgtaaagca agtacttaca tcaattggga agataactct gtcattttcc
     1321 tacttgttca gtccatggtg gttgatctta agaacctgct ttttaatcca agtaagccat
     1381 tctcaagagg cagtcagcct gcagatgtgg atctaatgat tgactgcctt gtttcttgct
     1441 ttcgtataag ccctcacaac aaccaacact ttaagatctg cctggctcag aattcacctt
     1501 ctacatttca ctatgtgctg gtaaattcac tccatcgaat catcaccaat tccgcattgg
     1561 attggtggcc taagattgat gctgtgtatt gtcactcggt tgaacttcga aatatgtttg
     1621 gtgaaacact tcataaagca gtgcaaggtt gtggagcaca cccagcaata cgaatggcac
     1681 cgagtcttac atttaaagaa aaagtaacaa gccttaaatt taaagaaaaa cctacagacc
     1741 tggagacaag aagctataag tatcttctct tgtccatggt gaaactaatt catgcagatc
     1801 caaagctctt gctttgtaat ccaagaaaac aggggcccga aacccaaggc agtacagcag
     1861 aattaattac agggctcgtc caactggtcc ctcagtcaca catgccagag attgctcagg
     1921 aagcaatgga ggctctgctg gttcttcatc agttagatag cattgatttg tggaatcctg
     1981 atgctcctgt agaaacattt tgggagatta gctcacaaat gcttttttac atctgcaaga
     2041 aattaactag tcatcaaatg cttagtagca cagaaattct caagtggttg cgggaaatat
     2101 tgatctgcag gaataaattt cttcttaaaa ataagcaggc agatagaagt tcctgtcact
     2161 ttctcctttt ttacggggta ggatgtgata ttccttctag tggaaatacc agtcaaatgt
     2221 ccatggatca tgaagaatta ctacgtactc ctggagcctc tctccggaag ggaaaaggga
     2281 actcctctat ggatagtgca gcaggatgca gcggaacccc cccaatttgc cgacaagccc
     2341 agaccaaact agaagtggcc ctgtacatgt ttctgtggaa ccctgacact gaagctgttc
     2401 tggttgccat gtcctgtttc cgccacctct gtgaggaagc agatatccgg tgtggggtgg
     2461 atgaagtgtc agtgcataac ctcttgccca actataacac attcatggag tttgcctctg
     2521 tcagcaatat gatgtcaaca ggaagagcag cacttcagaa aagagtgatg gcactgctga
     2581 ggcgcattga gcatcccact gcaggaaaca ctgaggcttg ggaagataca catgcaaaat
     2641 gggaacaagc aacaaagcta atccttaact atccaaaagc caaaatggaa gatggccagg
     2701 ctgctgaaag ccttcacaag accattgtca agaggcgaat gtcccatgtg agtggaggag
     2761 gatccataga tttgtctgac acagactccc tacaggaatg gatcaacatg actggcttcc
     2821 tttgtgccct tgggggagtg tgcctccagc agagaagcaa ttctggcctg gcaacctata
     2881 gcccacccat gggtccagtc agtgaacgta agggttctat gatttcagtg atgtcttcag
     2941 agggaaacgc agatacacct gtcagcaaat ttatggatcg gctgttgtcc ttaatggtgt
     3001 gtaaccatga gaaagtggga cttcaaatac ggaccaatgt taaggatctg gtgggtctag
     3061 aattgagtcc tgctctgtat ccaatgctat ttaacaaatt gaagaatacc atcagcaagt
     3121 tttttgactc ccaaggacag gttttattga ctgataccaa tactcaattt gtagaacaaa
     3181 ccatagctat aatgaagaac ttgctagata atcatactga aggcagctct gaacatctag
     3241 ggcaagctag cattgaaaca atgatgttaa atctggtcag gtatgttcgt gtgcttggga
     3301 atatggtcca tgcaattcaa ataaaaacga aactgtgtca attagttgaa gtaatgatgg
     3361 caaggagaga tgacctctca ttttgccaag agatgaaatt taggaataag atggtagaat
     3421 acctgacaga ctgggttatg ggaacatcaa accaagcagc agatgatgat gtaaaatgtc
     3481 ttacaagaga tttggaccag gcaagcatgg aagcagtagt ttcacttcta gctggtctcc
     3541 ctctgcagcc tgaagaagga gatggtgtgg aattgatgga agccaaatca cagttatttc
     3601 ttaaatactt cacattattt atgaaccttt tgaatgactg cagtgaagtt gaagatgaaa
     3661 gtgcgcaaac aggtggcagg aaacgtggca tgtctcggag gctggcatca ctgaggcact
     3721 gtacggtcct tgcaatgtca aacttactca atgccaacgt agacagtggt ctcatgcact
     3781 ccataggctt aggttaccac aaggatctcc agacaagagc tacatttatg gaagttctga
     3841 caaaaatcct tcaacaaggc acagaatttg acacacttgc agaaacagta ttggctgatc
     3901 ggtttgagag attggtggaa ctggtcacaa tgatgggtga tcaaggagaa ctccctatag
     3961 cgatggctct ggccaatgtg gttccttgtt ctcagtggga tgaactagct cgagttctgg
     4021 ttactctgtt tgattctcgg catttactct accaactgct ctggaacatg ttttctaaag
     4081 aagtagaatt ggcagactcc atgcagactc tcttccgagg caacagcttg gccagtaaaa
     4141 taatgacatt ctgtttcaag gtatatggtg ctacctatct acaaaaactc ctggatcctt
     4201 tattacgaat tgtgatcaca tcctctgatt ggcaacatgt tagctttgaa gtggatccta
     4261 ccaggttaga accatcagag agccttgagg aaaaccagcg gaacctcctt cagatgactg
     4321 aaaagttctt ccatgccatc atcagttcct cctcagaatt cccccctcaa cttcgaagtg
     4381 tgtgccactg tttataccag gcaacttgcc actccctact gaataaagct acagtaaaag
     4441 aaaaaaagga aaacaaaaaa tcagtggtta gccagcgttt ccctcagaac agcatcggtg
     4501 cagtaggaag tgccatgttc ctcagattta tcaatcctgc cattgtctca ccgtatgaag
     4561 cagggatttt agataaaaag ccaccaccta gaatcgaaag gggcttgaag ttaatgtcaa
     4621 agatacttca gagtattgcc aatcatgttc tcttcacaaa agaagaacat atgcggcctt
     4681 tcaatgattt tgtgaaaagc aactttgatg cagcacgcag gtttttcctt gatatagcat
     4741 ctgattgtcc tacaagtgat gcagtaaatc atagtctttc cttcataagt gacggcaatg
     4801 tgcttgcttt acatcgtcta ctctggaaca atcaggagaa aattgggcag tatctttcca
     4861 gcaacaggga tcataaagct gttggaagac gaccttttga taagatggca acacttcttg
     4921 catacctggg tcctccagag cacaaacctg tggcagatac acactggtcc agccttaacc
     4981 ttaccagttc aaagtttgag gaatttatga ctaggcatca ggtacatgaa aaagaagaat
     5041 tcaaggcttt gaaaacgtta agtattttct accaagctgg gacttccaaa gctgggaatc
     5101 ctatttttta ttatgttgca cggaggtaag aaatactatg ttttgggtct cttaacagaa
     5161 ttttttaaat tatagcaaat atagagagat ggcaactaga ttagtggcat ctgtatattt
     5221 attttaaaca ctgctaataa tctttgtctt ttttgtcatt ttccttaggt tcaaaactgg
     5281 tcaaatcaat ggtgatttgc tgatatacca tgtcttactg actttaaagc catattatgc
     5341 aaagccatat gaaattgtag tggaccttac ccataccggg cctagcaatc gctttaaaac
     5401 agactttctc tctaagtggt ttgttgtttt tcctggcttt gcttacgaca acgtctccgc
     5461 agtctatatc tataactgta actcctgggt cagggagtac accaagtatc atgagcggct
     5521 gctgactggc ctcaaaggta gcaaaaggct tgttttcata gactgtcctg ggaaactggc
     5581 tgagcacata gagcatgaac aacagaaact acctgctgcc accttggctt tagaagagga
     5641 cctgaaggta ttccacaatg ctctcaagct agctcacaaa gacaccaaag tttctattaa
     5701 agttggttct actgctgtcc aagtaacttc agcagagcga acaaaagtcc tagggcaatc
     5761 agtctttcta aatgacattt attatgcttc ggaaattgaa gaaatctgcc tagtagatga
     5821 gaaccagttc accttaacca ttgcaaacca gggcacgccg ctcaccttca tgcaccagga
     5881 gtgtgaagcc attgtccagt ctatcattca tatccggacc cgctgggaac tgtcacagcc
     5941 cgactctatc ccccaacaca ccaagattcg gccaaaagat gtccctggga cactgctcaa
     6001 tatcgcatta cttaatttag gcagttctga cccgagttta cggtcagctg cctataatct
     6061 tctgtgtgcc ttaacttgta cctttaattt aaaaatcgag ggccagttac tagagacatc
     6121 aggtttatgt atccctgcca acaacaccct ctttattgtc tctattagta agacactggc
     6181 agccaatgag ccacacctca cgttagaatt tttggaagag tgtatttctg gatttagcaa
     6241 atctagtatt gaattgaaac acctttgttt ggaatacatg actccatggc tgtcaaatct
     6301 agttcgtttt tgcaagcata atgatgatgc caaacgacaa agagttactg ctattcttga
     6361 caagctgata acaatgacca tcaatgaaaa acagatgtac ccatctattc aagcaaaaat
     6421 atggggaagc cttgggcaga ttacagatct gcttgatgtt gtactagaca gtttcatcaa
     6481 aaccagtgca acaggtggct tgggatcaat aaaagctgag gtgatggcag atactgctgt
     6541 agctttggct tctggaaatg tgaaattggt ttcaagcaag gttattggaa ggatgtgcaa
     6601 aataattgac aagacatgct tatctccaac tcctacttta gaacaacatc ttatgtggga
     6661 tgatattgct attttagcac gctacatgct gatgctgtcc ttcaacaatt cccttgatgt
     6721 ggcagctcat cttccctacc tcttccacgt tgttactttc ttagtagcca caggtccgct
     6781 ctcccttaga gcttccacac atggactggt cattaatatc attcactctc tgtgtacttg
     6841 ttcacagctt cattttagtg aagagaccaa gcaagttttg agactcagtc tgacagagtt
     6901 ctcattaccc aaattttact tgctgtttgg cattagcaaa gtcaagtcag ctgctgtcat
     6961 tgccttccgt tccagttacc gggacaggtc attctctcct ggctcctatg agagagagac
     7021 ttttgctttg acatccttgg aaacagtcac agaagctttg ttggagatca tggaggcatg
     7081 catgagagat attccaacgt gcaagtggct ggaccagtgg acagaactag ctcaaagatt
     7141 tgcattccaa tataatccat ccctgcaacc aagagctctt gttgtctttg ggtgtattag
     7201 caaacgagtg tctcatgggc agataaagca gataatccgt attcttagca aggcacttga
     7261 gagttgctta aaaggacctg acacttacaa cagtcaagtt ctgatagaag ctacagtaat
     7321 agcactaacc aaattacagc cacttcttaa taaggactcg cctctgcaca aagccctctt
     7381 ttgggtagct gtggctgtgc tgcagcttga tgaggtcaac ttgtattcag caggtaccgc
     7441 acttcttgaa caaaacctgc atactttaga tagtctccgt atattcaatg acaagagtcc
     7501 agaggaagta tttatggcaa tccggaatcc tctggagtgg cactgcaagc aaatggatca
     7561 ttttgttgga ctcaatttca actctaactt taactttgca ttggttggac accttttaaa
     7621 agggtacagg catccttcac ctgctattgt tgcaagaaca gtcagaattt tacatacact
     7681 actaactctg gttaacaaac acagaaattg tgacaaattt gaagtgaata cacagagcgt
     7741 ggcctactta gcagctttac ttacagtgtc tgaagaagtt cgaagtcgct gcagcctaaa
     7801 acatagaaag tcacttcttc ttactgatat ttcaatggaa aatgttccta tggatacata
     7861 tcccattcat catggtgacc cttcctatag gacactaaag gagactcagc catggtcctc
     7921 tcccaaaggt tctgaaggat accttgcagc cacctatcca actgtcggcc agaccagtcc
     7981 ccgagccagg aaatccatga gcctggacat ggggcaacct tctcaggcca acactaagaa
     8041 gttgcttgga acaaggaaaa gttttgatca cttgatatca gacacaaagg ctcctaaaag
     8101 gcaagaaatg gaatcaggga tcacaacacc ccccaaaatg aggagagtag cagaaactga
     8161 ttatgaaatg gaaactcaga ggatttcctc atcacaacag cacccacatt tacgtaaagt
     8221 ttcagtgtct gaatcaaatg ttctcttgga tgaagaagta cttactgatc cgaagatcca
     8281 ggcgctgctt cttactgttc tagctacact ggtaaaatat accacagatg agtttgatca
     8341 acgaattctt tatgaatact tagcagaggc cagtgttgtg tttcccaaag tctttcctgt
     8401 tgtgcataat ttgttggact ctaagatcaa caccctgtta tcattgtgcc aagatccaaa
     8461 tttgttaaat ccaatccatg gaattgtgca gagtgtggtg taccatgaag aatccccacc
     8521 acaataccaa acatcttacc tgcaaagttt tggttttaat ggcttgtggc ggtttgcagg
     8581 accgttttca aagcaaacac aaattccaga ctatgctgag cttattgtta agtttcttga
     8641 tgccttgatt gacacgtacc tgcctggaat tgatgaagaa accagtgaag aatccctcct
     8701 gactcccaca tctccttacc ctcctgcact gcagagccag cttagtatca ctgccaacct
     8761 taacctttct aattccatga cctcacttgc aacttcccag cattccccag gaatcgacaa
     8821 ggagaacgtt gaactctccc ctaccactgg ccactgtaac agtggacgaa ctcgccacgg
     8881 atccgcaagc caagtgcaga agcaaagaag cgcaggatca tttaagagaa atagcattaa
     8941 gaagatcgtg tagcggccgc atcgtgactg actgacgatc tgcctcgcgc gtttcggtga
     9001 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     9061 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     9121 cgcagccatg acccagtcac gtagcgatag cggagtgtat aattcttgaa gacgaaaggg
     9181 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     9241 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     9301 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     9361 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     9421 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     9481 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     9541 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     9601 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     9661 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     9721 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     9781 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     9841 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     9901 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     9961 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
    10021 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
    10081 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
    10141 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
    10201 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
    10261 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
    10321 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
    10381 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
    10441 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
    10501 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
    10561 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
    10621 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
    10681 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
    10741 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcg
//
