LOCUS       sequence-229908-       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 R1276P (+minintr, KDR)
FEATURES             Location/Qualifiers
     source          1..10788
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(442..464)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(609..627)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        695..799
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(800..868,869..1660)
                     /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(1018..1037)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1831..2419
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2320..2339
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtttgcagga ccgttttcaa agcaaacaca aattccagac tatgctgagc ttattgttaa
       61 gtttcttgat gccttgattg acacgtacct gcctggaatt gatgaagaaa ccagtgaaga
      121 atccctcctg actcccacat ctccttaccc tcctgcactg cagagccagc ttagtatcac
      181 tgccaacctt aacctttcta attccatgac ctcacttgca acttcccagc attccccagg
      241 aatcgacaag gagaacgttg aactctcccc taccactggc cactgtaaca gtggacgaac
      301 tcgccacgga tccgcaagcc aagtgcagaa gcaaagaagc gcaggatcat ttaagagaaa
      361 tagcattaag aagatcgtgt agcggccgca tcgtgactga ctgacgatct gcctcgcgcg
      421 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
      481 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
      541 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata attcttgaag
      601 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
      661 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
      721 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
      781 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
      841 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
      901 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
      961 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     1021 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     1081 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     1141 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     1201 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     1261 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     1321 cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg
     1381 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     1441 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     1501 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     1561 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     1621 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     1681 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     1741 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     1801 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     1861 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     1921 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     1981 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2041 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2101 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2161 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2221 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2281 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2341 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2401 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2461 ctggcctttt gctcacatgt cgtacgacgc gtcccgggac cggtggcatg gccgcgcaca
     2521 ggccggtgga atgggtccag gccgtggtca gccgcttcga cgagcagctt ccaataaaaa
     2581 caggacagca gaacacacat accaaagtca gtactgagca caacaaggaa tgtctaatca
     2641 atatttccaa atacaagttt tctttggtta taagcggcct cactactatt ttaaagaatg
     2701 ttaacaatat gagaatattt ggagaagctg ctgaaaaaaa tttatatctc tctcagttga
     2761 ttatattgga tacactggaa aaatgtcttg ctgggcaacc aaaggacaca atgagattag
     2821 atgaaacgat gctggtcaaa cagttgctgc cagaaatctg ccattttctt cacacctgtc
     2881 gtgaaggaaa ccagcatgca gctgaacttc ggaattctgc ctctggggtt ttattttctc
     2941 tcagctgcaa caacttcaat gcagtcttta gtcgcatttc taccaggtta caggaattaa
     3001 ctgtttgttc agaagacaat gttgatgttc atgatataga attgttacag tatatcaatg
     3061 tggattgtgc aaaattaaaa cgactcctga aggaaacagc atttaaattt aaagccctaa
     3121 agaaggttgc gcagttagca gttataaata gcctggaaaa ggcattttgg aactgggtag
     3181 aaaattatcc agatgaattt acaaaactat accagatccc acagactgat atggctgaat
     3241 gtgcagaaaa gctatttgac ttggtggatg gttttgctga aagcaccaaa cgtaaagcag
     3301 cagtttggcc actacaaatc attctcctta tcttgtgtcc agaaataatc caggatatat
     3361 ccaaagacgt ggttgatgaa aacaacatga ataagaagtt atttctggac agtctacgaa
     3421 aagctcttgc tggccatgga ggaagtaggc agctgacaga aagtgctgca attgcctgtg
     3481 tcaaactgtg taaagcaagt acttacatca attgggaaga taactctgtc attttcctac
     3541 ttgttcagtc catggtggtt gatcttaaga acctgctttt taatccaagt aagccattct
     3601 caagaggcag tcagcctgca gatgtggatc taatgattga ctgccttgtt tcttgctttc
     3661 gtataagccc tcacaacaac caacacttta agatctgcct ggctcagaat tcaccttcta
     3721 catttcacta tgtgctggta aattcactcc atcgaatcat caccaattcc gcattggatt
     3781 ggtggcctaa gattgatgct gtgtattgtc actcggttga acttcgaaat atgtttggtg
     3841 aaacacttca taaagcagtg caaggttgtg gagcacaccc agcaatacga atggcaccga
     3901 gtcttacatt taaagaaaaa gtaacaagcc ttaaatttaa agaaaaacct acagacctgg
     3961 agacaagaag ctataagtat cttctcttgt ccatggtgaa actaattcat gcagatccaa
     4021 agctcttgct ttgtaatcca agaaaacagg ggcccgaaac ccaaggcagt acagcagaat
     4081 taattacagg gctcgtccaa ctggtccctc agtcacacat gccagagatt gctcaggaag
     4141 caatggaggc tctgctggtt cttcatcagt tagatagcat tgatttgtgg aatcctgatg
     4201 ctcctgtaga aacattttgg gagattagct cacaaatgct tttttacatc tgcaagaaat
     4261 taactagtca tcaaatgctt agtagcacag aaattctcaa gtggttgcgg gaaatattga
     4321 tctgcaggaa taaatttctt cttaaaaata agcaggcaga tagaagttcc tgtcactttc
     4381 tcctttttta cggggtagga tgtgatattc cttctagtgg aaataccagt caaatgtcca
     4441 tggatcatga agaattacta cgtactcctg gagcctctct ccggaaggga aaagggaact
     4501 cctctatgga tagtgcagca ggatgcagcg gaaccccccc aatttgccga caagcccaga
     4561 ccaaactaga agtggccctg tacatgtttc tgtggaaccc tgacactgaa gctgttctgg
     4621 ttgccatgtc ctgtttccgc cacctctgtg aggaagcaga tatccggtgt ggggtggatg
     4681 aagtgtcagt gcataacctc ttgcccaact ataacacatt catggagttt gcctctgtca
     4741 gcaatatgat gtcaacagga agagcagcac ttcagaaaag agtgatggca ctgctgaggc
     4801 gcattgagca tcccactgca ggaaacactg aggcttggga agatacacat gcaaaatggg
     4861 aacaagcaac aaagctaatc cttaactatc caaaagccaa aatggaagat ggccaggctg
     4921 ctgaaagcct tcacaagacc attgtcaaga ggcgaatgtc ccatgtgagt ggaggaggat
     4981 ccatagattt gtctgacaca gactccctac aggaatggat caacatgact ggcttccttt
     5041 gtgcccttgg gggagtgtgc ctccagcaga gaagcaattc tggcctggca acctatagcc
     5101 cacccatggg tccagtcagt gaacgtaagg gttctatgat ttcagtgatg tcttcagagg
     5161 gaaacgcaga tacacctgtc agcaaattta tggatcggct gttgtcctta atggtgtgta
     5221 accatgagaa agtgggactt caaatacgga ccaatgttaa ggatctggtg ggtctagaat
     5281 tgagtcctgc tctgtatcca atgctattta acaaattgaa gaataccatc agcaagtttt
     5341 ttgactccca aggacaggtt ttattgactg ataccaatac tcaatttgta gaacaaacca
     5401 tagctataat gaagaacttg ctagataatc atactgaagg cagctctgaa catctagggc
     5461 aagctagcat tgaaacaatg atgttaaatc tggtcaggta tgttcgtgtg cttgggaata
     5521 tggtccatgc aattcaaata aaaacgaaac tgtgtcaatt agttgaagta atgatggcaa
     5581 ggagagatga cctctcattt tgccaagaga tgaaatttag gaataagatg gtagaatacc
     5641 tgacagactg ggttatggga acatcaaacc aagcagcaga tgatgatgta aaatgtctta
     5701 caagagattt ggaccaggca agcatggaag cagtagtttc acttctagct ggtctccctc
     5761 tgcagcctga agaaggagat ggtgtggaat tgatggaagc caaatcacag ttatttctta
     5821 aatacttcac attatttatg aaccttttga atgactgcag tgaagttgaa gatgaaagtg
     5881 cgcaaacagg tggcaggaaa cgtggcatgt ctcggaggct ggcatcactg aggcactgta
     5941 cggtccttgc aatgtcaaac ttactcaatg ccaacgtaga cagtggtctc atgcactcca
     6001 taggcttagg ttaccacaag gatctccaga caagagctac atttatggaa gttctgacaa
     6061 aaatccttca acaaggcaca gaatttgaca cacttgcaga aacagtattg gctgatcggt
     6121 ttgagagatt ggtggaactg gtcacaatga tgggtgatca aggagaactc cctatagcga
     6181 tggctctggc caatgtggtt ccttgttctc agtgggatga actagctcga gttctggtta
     6241 ctctgtttga ttctcggcat ttactctacc aactgctctg gaacatgttt tctaaagaag
     6301 tagaattggc agactccatg cagactctct tcccaggcaa cagcttggcc agtaaaataa
     6361 tgacattctg tttcaaggta tatggtgcta cctatctaca aaaactcctg gatcctttat
     6421 tacgaattgt gatcacatcc tctgattggc aacatgttag ctttgaagtg gatcctacca
     6481 ggttagaacc atcagagagc cttgaggaaa accagcggaa cctccttcag atgactgaaa
     6541 agttcttcca tgccatcatc agttcctcct cagaattccc ccctcaactt cgaagtgtgt
     6601 gccactgttt ataccaggca acttgccact ccctactgaa taaagctaca gtaaaagaaa
     6661 aaaaggaaaa caaaaaatca gtggttagcc agcgtttccc tcagaacagc atcggtgcag
     6721 taggaagtgc catgttcctc agatttatca atcctgccat tgtctcaccg tatgaagcag
     6781 ggattttaga taaaaagcca ccacctagaa tcgaaagggg cttgaagtta atgtcaaaga
     6841 tacttcagag tattgccaat catgttctct tcacaaaaga agaacatatg cggcctttca
     6901 atgattttgt gaaaagcaac tttgatgcag cacgcaggtt tttccttgat atagcatctg
     6961 attgtcctac aagtgatgca gtaaatcata gtctttcctt cataagtgac ggcaatgtgc
     7021 ttgctttaca tcgtctactc tggaacaatc aggagaaaat tgggcagtat ctttccagca
     7081 acagggatca taaagctgtt ggaagacgac cttttgataa gatggcaaca cttcttgcat
     7141 acctgggtcc tccagagcac aaacctgtgg cagatacaca ctggtccagc cttaacctta
     7201 ccagttcaaa gtttgaggaa tttatgacta ggcatcaggt acatgaaaaa gaagaattca
     7261 aggctttgaa aacgttaagt attttctacc aagctgggac ttccaaagct gggaatccta
     7321 ttttttatta tgttgcacgg aggtaagaaa tactatgttt tgggtctctt aacagaattt
     7381 tttaaattat agcaaatata gagagatggc aactagatta gtggcatctg tatatttatt
     7441 ttaaacactg ctaataatct ttgtcttttt tgtcattttc cttaggttca aaactggtca
     7501 aatcaatggt gatttgctga tataccatgt cttactgact ttaaagccat attatgcaaa
     7561 gccatatgaa attgtagtgg accttaccca taccgggcct agcaatcgct ttaaaacaga
     7621 ctttctctct aagtggtttg ttgtttttcc tggctttgct tacgacaacg tctccgcagt
     7681 ctatatctat aactgtaact cctgggtcag ggagtacacc aagtatcatg agcggctgct
     7741 gactggcctc aaaggtagca aaaggcttgt tttcatagac tgtcctggga aactggctga
     7801 gcacatagag catgaacaac agaaactacc tgctgccacc ttggctttag aagaggacct
     7861 gaaggtattc cacaatgctc tcaagctagc tcacaaagac accaaagttt ctattaaagt
     7921 tggttctact gctgtccaag taacttcagc agagcgaaca aaagtcctag ggcaatcagt
     7981 ctttctaaat gacatttatt atgcttcgga aattgaagaa atctgcctag tagatgagaa
     8041 ccagttcacc ttaaccattg caaaccaggg cacgccgctc accttcatgc accaggagtg
     8101 tgaagccatt gtccagtcta tcattcatat ccggacccgc tgggaactgt cacagcccga
     8161 ctctatcccc caacacacca agattcggcc aaaagatgtc cctgggacac tgctcaatat
     8221 cgcattactt aatttaggca gttctgaccc gagtttacgg tcagctgcct ataatcttct
     8281 gtgtgcctta acttgtacct ttaatttaaa aatcgagggc cagttactag agacatcagg
     8341 tttatgtatc cctgccaaca acaccctctt tattgtctct attagtaaga cactggcagc
     8401 caatgagcca cacctcacgt tagaattttt ggaagagtgt atttctggat ttagcaaatc
     8461 tagtattgaa ttgaaacacc tttgtttgga atacatgact ccatggctgt caaatctagt
     8521 tcgtttttgc aagcataatg atgatgccaa acgacaaaga gttactgcta ttcttgacaa
     8581 gctgataaca atgaccatca atgaaaaaca gatgtaccca tctattcaag caaaaatatg
     8641 gggaagcctt gggcagatta cagatctgct tgatgttgta ctagacagtt tcatcaaaac
     8701 cagtgcaaca ggtggcttgg gatcaataaa agctgaggtg atggcagata ctgctgtagc
     8761 tttggcttct ggaaatgtga aattggtttc aagcaaggtt attggaagga tgtgcaaaat
     8821 aattgacaag acatgcttat ctccaactcc tactttagaa caacatctta tgtgggatga
     8881 tattgctatt ttagcacgct acatgctgat gctgtccttc aacaattccc ttgatgtggc
     8941 agctcatctt ccctacctct tccacgttgt tactttctta gtagccacag gtccgctctc
     9001 ccttagagct tccacacatg gactggtcat taatatcatt cactctctgt gtacttgttc
     9061 acagcttcat tttagtgaag agaccaagca agttttgaga ctcagtctga cagagttctc
     9121 attacccaaa ttttacttgc tgtttggcat tagcaaagtc aagtcagctg ctgtcattgc
     9181 cttccgttcc agttaccggg acaggtcatt ctctcctggc tcctatgaga gagagacttt
     9241 tgctttgaca tccttggaaa cagtcacaga agctttgttg gagatcatgg aggcatgcat
     9301 gagagatatt ccaacgtgca agtggctgga ccagtggaca gaactagctc aaagatttgc
     9361 attccaatat aatccatccc tgcaaccaag agctcttgtt gtctttgggt gtattagcaa
     9421 acgagtgtct catgggcaga taaagcagat aatccgtatt cttagcaagg cacttgagag
     9481 ttgcttaaaa ggacctgaca cttacaacag tcaagttctg atagaagcta cagtaatagc
     9541 actaaccaaa ttacagccac ttcttaataa ggactcgcct ctgcacaaag ccctcttttg
     9601 ggtagctgtg gctgtgctgc agcttgatga ggtcaacttg tattcagcag gtaccgcact
     9661 tcttgaacaa aacctgcata ctttagatag tctccgtata ttcaatgaca agagtccaga
     9721 ggaagtattt atggcaatcc ggaatcctct ggagtggcac tgcaagcaaa tggatcattt
     9781 tgttggactc aatttcaact ctaactttaa ctttgcattg gttggacacc ttttaaaagg
     9841 gtacaggcat ccttcacctg ctattgttgc aagaacagtc agaattttac atacactact
     9901 aactctggtt aacaaacaca gaaattgtga caaatttgaa gtgaatacac agagcgtggc
     9961 ctacttagca gctttactta cagtgtctga agaagttcga agtcgctgca gcctaaaaca
    10021 tagaaagtca cttcttctta ctgatatttc aatggaaaat gttcctatgg atacatatcc
    10081 cattcatcat ggtgaccctt cctataggac actaaaggag actcagccat ggtcctctcc
    10141 caaaggttct gaaggatacc ttgcagccac ctatccaact gtcggccaga ccagtccccg
    10201 agccaggaaa tccatgagcc tggacatggg gcaaccttct caggccaaca ctaagaagtt
    10261 gcttggaaca aggaaaagtt ttgatcactt gatatcagac acaaaggctc ctaaaaggca
    10321 agaaatggaa tcagggatca caacaccccc caaaatgagg agagtagcag aaactgatta
    10381 tgaaatggaa actcagagga tttcctcatc acaacagcac ccacatttac gtaaagtttc
    10441 agtgtctgaa tcaaatgttc tcttggatga agaagtactt actgatccga agatccaggc
    10501 gctgcttctt actgttctag ctacactggt aaaatatacc acagatgagt ttgatcaacg
    10561 aattctttat gaatacttag cagaggccag tgttgtgttt cccaaagtct ttcctgttgt
    10621 gcataatttg ttggactcta agatcaacac cctgttatca ttgtgccaag atccaaattt
    10681 gttaaatcca atccatggaa ttgtgcagag tgtggtgtac catgaagaat ccccaccaca
    10741 ataccaaaca tcttacctgc aaagttttgg ttttaatggc ttgtggcg
//
