LOCUS       sequence-225451-       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"
     rep_origin      79..667
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     568..587
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(9478..9500)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9645..9663)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9731..9835
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9836..9904,9905..10696)
                     /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(10054..10073)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
       61 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
      121 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
      181 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
      241 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      301 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
      361 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      421 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      481 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
      541 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      601 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      661 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
      721 tcacatgtcg tacgacgcgt cccgggaccg gtggcatggc cgcgcacagg ccggtggaat
      781 gggtccaggc cgtggtcagc cgcttcgacg agcagcttcc aataaaaaca ggacagcaga
      841 acacacatac caaagtcagt actgagcaca acaaggaatg tctaatcaat atttccaaat
      901 acaagttttc tttggttata agcggcctca ctactatttt aaagaatgtt aacaatatga
      961 gaatatttgg agaagctgct gaaaaaaatt tatatctctc tcagttgatt atattggata
     1021 cactggaaaa atgtcttgct gggcaaccaa aggacacaat gagattagat gaaacgatgc
     1081 tggtcaaaca gttgctgcca gaaatctgcc attttcttca cacctgtcgt gaaggaaacc
     1141 agcatgcagc tgaacttcgg aattctgcct ctggggtttt attttctctc agctgcaaca
     1201 acttcaatgc agtctttagt cgcatttcta ccaggttaca ggaattaact gtttgttcag
     1261 aagacaatgt tgatgttcat gatatagaat tgttacagta tatcaatgtg gattgtgcaa
     1321 aattaaaacg actcctgaag gaaacagcat ttaaatttaa agccctaaag aaggttgcgc
     1381 agttagcagt tataaatagc ctggaaaagg cattttggaa ctgggtagaa aattatccag
     1441 atgaatttac aaaactatac cagatcccac agactgatat ggctgaatgt gcagaaaagc
     1501 tatttgactt ggtggatggt tttgctgaaa gcaccaaacg taaagcagca gtttggccac
     1561 tacaaatcat tctccttatc ttgtgtccag aaataatcca ggatatatcc aaagacgtgg
     1621 ttgatgaaaa caacatgaat aagaagttat ttctggacag tctacgaaaa gctcttgctg
     1681 gccatggagg aagtaggcag ctgacagaaa gtgctgcaat tgcctgtgtc aaactgtgta
     1741 aagcaagtac ttacatcaat tgggaagata actctgtcat tttcctactt gttcagtcca
     1801 tggtggttga tcttaagaac ctgcttttta atccaagtaa gccattctca agaggcagtc
     1861 agcctgcaga tgtggatcta atgattgact gccttgtttc ttgctttcgt ataagccctc
     1921 acaacaacca acactttaag atctgcctgg ctcagaattc accttctaca tttcactatg
     1981 tgctggtaaa ttcactccat cgaatcatca ccaattccgc attggattgg tggcctaaga
     2041 ttgatgctgt gtattgtcac tcggttgaac ttcgaaatat gtttggtgaa acacttcata
     2101 aagcagtgca aggttgtgga gcacacccag caatacgaat ggcaccgagt cttacattta
     2161 aagaaaaagt aacaagcctt aaatttaaag aaaaacctac agacctggag acaagaagct
     2221 ataagtatct tctcttgtcc atggtgaaac taattcatgc agatccaaag ctcttgcttt
     2281 gtaatccaag aaaacagggg cccgaaaccc aaggcagtac agcagaatta attacagggc
     2341 tcgtccaact ggtccctcag tcacacatgc cagagattgc tcaggaagca atggaggctc
     2401 tgctggttct tcatcagtta gatagcattg atttgtggaa tcctgatgct cctgtagaaa
     2461 cattttggga gattagctca caaatgcttt tttacatctg caagaaatta actagtcatc
     2521 aaatgcttag tagcacagaa attctcaagt ggttgcggga aatattgatc tgcaggaata
     2581 aatttcttct taaaaataag caggcagata gaagttcctg tcactttctc cttttttacg
     2641 gggtaggatg tgatattcct tctagtggaa ataccagtca aatgtccatg gatcatgaag
     2701 aattactacg tactcctgga gcctctctcc ggaagggaaa agggaactcc tctatggata
     2761 gtgcagcagg atgcagcgga acccccccaa tttgccgaca agcccagacc aaactagaag
     2821 tggccctgta catgtttctg tggaaccctg acactgaagc tgttctggtt gccatgtcct
     2881 gtttccgcca cctctgtgag gaagcagata tccggtgtgg ggtggatgaa gtgtcagtgc
     2941 ataacctctt gcccaactat aacacattca tggagtttgc ctctgtcagc aatatgatgt
     3001 caacaggaag agcagcactt cagaaaagag tgatggcact gctgaggcgc attgagcatc
     3061 ccactgcagg aaacactgag gcttgggaag atacacatgc aaaatgggaa caagcaacaa
     3121 agctaatcct taactatcca aaagccaaaa tggaagatgg ccaggctgct gaaagccttc
     3181 acaagaccat tgtcaagagg cgaatgtccc atgtgagtgg aggaggatcc atagatttgt
     3241 ctgacacaga ctccctacag gaatggatca acatgactgg cttcctttgt gcccttgggg
     3301 gagtgtgcct ccagcagaga agcaattctg gcctggcaac ctatagccca cccatgggtc
     3361 cagtcagtga acgtaagggt tctatgattt cagtgatgtc ttcagaggga aacgcagata
     3421 cacctgtcag caaatttatg gatcggctgt tgtccttaat ggtgtgtaac catgagaaag
     3481 tgggacttca aatacggacc aatgttaagg atctggtggg tctagaattg agtcctgctc
     3541 tgtatccaat gctatttaac aaattgaaga ataccatcag caagtttttt gactcccaag
     3601 gacaggtttt attgactgat accaatactc aatttgtaga acaaaccata gctataatga
     3661 agaacttgct agataatcat actgaaggca gctctgaaca tctagggcaa gctagcattg
     3721 aaacaatgat gttaaatctg gtcaggtatg ttcgtgtgct tgggaatatg gtccatgcaa
     3781 ttcaaataaa aacgaaactg tgtcaattag ttgaagtaat gatggcaagg agagatgacc
     3841 tctcattttg ccaagagatg aaatttagga ataagatggt agaatacctg acagactggg
     3901 ttatgggaac atcaaaccaa gcagcagatg atgatgtaaa atgtcttaca agagatttgg
     3961 accaggcaag catggaagca gtagtttcac ttctagctgg tctccctctg cagcctgaag
     4021 aaggagatgg tgtggaattg atggaagcca aatcacagtt atttcttaaa tacttcacat
     4081 tatttatgaa ccttttgaat gactgcagtg aagttgaaga tgaaagtgcg caaacaggtg
     4141 gcaggaaacg tggcatgtct cggaggctgg catcactgag gcactgtacg gtccttgcaa
     4201 tgtcaaactt actcaatgcc aacgtagaca gtggtctcat gcactccata ggcttaggtt
     4261 accacaagga tctccagaca agagctacat ttatggaagt tctgacaaaa atccttcaac
     4321 aaggcacaga atttgacaca cttgcagaaa cagtattggc tgatcggttt gagagattgg
     4381 tggaactggt cacaatgatg ggtgatcaag gagaactccc tatagcgatg gctctggcca
     4441 atgtggttcc ttgttctcag tgggatgaac tagctcgagt tctggttact ctgtttgatt
     4501 ctcggcattt actctaccaa ctgctctgga acatgttttc taaagaagta gaattggcag
     4561 actccatgca gactctcttc ccaggcaaca gcttggccag taaaataatg acattctgtt
     4621 tcaaggtata tggtgctacc tatctacaaa aactcctgga tcctttatta cgaattgtga
     4681 tcacatcctc tgattggcaa catgttagct ttgaagtgga tcctaccagg ttagaaccat
     4741 cagagagcct tgaggaaaac cagcggaacc tccttcagat gactgaaaag ttcttccatg
     4801 ccatcatcag ttcctcctca gaattccccc ctcaacttcg aagtgtgtgc cactgtttat
     4861 accaggcaac ttgccactcc ctactgaata aagctacagt aaaagaaaaa aaggaaaaca
     4921 aaaaatcagt ggttagccag cgtttccctc agaacagcat cggtgcagta ggaagtgcca
     4981 tgttcctcag atttatcaat cctgccattg tctcaccgta tgaagcaggg attttagata
     5041 aaaagccacc acctagaatc gaaaggggct tgaagttaat gtcaaagata cttcagagta
     5101 ttgccaatca tgttctcttc acaaaagaag aacatatgcg gcctttcaat gattttgtga
     5161 aaagcaactt tgatgcagca cgcaggtttt tccttgatat agcatctgat tgtcctacaa
     5221 gtgatgcagt aaatcatagt ctttccttca taagtgacgg caatgtgctt gctttacatc
     5281 gtctactctg gaacaatcag gagaaaattg ggcagtatct ttccagcaac agggatcata
     5341 aagctgttgg aagacgacct tttgataaga tggcaacact tcttgcatac ctgggtcctc
     5401 cagagcacaa acctgtggca gatacacact ggtccagcct taaccttacc agttcaaagt
     5461 ttgaggaatt tatgactagg catcaggtac atgaaaaaga agaattcaag gctttgaaaa
     5521 cgttaagtat tttctaccaa gctgggactt ccaaagctgg gaatcctatt ttttattatg
     5581 ttgcacggag gtaagaaata ctatgttttg ggtctcttaa cagaattttt taaattatag
     5641 caaatataga gagatggcaa ctagattagt ggcatctgta tatttatttt aaacactgct
     5701 aataatcttt gtcttttttg tcattttcct taggttcaaa actggtcaaa tcaatggtga
     5761 tttgctgata taccatgtct tactgacttt aaagccatat tatgcaaagc catatgaaat
     5821 tgtagtggac cttacccata ccgggcctag caatcgcttt aaaacagact ttctctctaa
     5881 gtggtttgtt gtttttcctg gctttgctta cgacaacgtc tccgcagtct atatctataa
     5941 ctgtaactcc tgggtcaggg agtacaccaa gtatcatgag cggctgctga ctggcctcaa
     6001 aggtagcaaa aggcttgttt tcatagactg tcctgggaaa ctggctgagc acatagagca
     6061 tgaacaacag aaactacctg ctgccacctt ggctttagaa gaggacctga aggtattcca
     6121 caatgctctc aagctagctc acaaagacac caaagtttct attaaagttg gttctactgc
     6181 tgtccaagta acttcagcag agcgaacaaa agtcctaggg caatcagtct ttctaaatga
     6241 catttattat gcttcggaaa ttgaagaaat ctgcctagta gatgagaacc agttcacctt
     6301 aaccattgca aaccagggca cgccgctcac cttcatgcac caggagtgtg aagccattgt
     6361 ccagtctatc attcatatcc ggacccgctg ggaactgtca cagcccgact ctatccccca
     6421 acacaccaag attcggccaa aagatgtccc tgggacactg ctcaatatcg cattacttaa
     6481 tttaggcagt tctgacccga gtttacggtc agctgcctat aatcttctgt gtgccttaac
     6541 ttgtaccttt aatttaaaaa tcgagggcca gttactagag acatcaggtt tatgtatccc
     6601 tgccaacaac accctcttta ttgtctctat tagtaagaca ctggcagcca atgagccaca
     6661 cctcacgtta gaatttttgg aagagtgtat ttctggattt agcaaatcta gtattgaatt
     6721 gaaacacctt tgtttggaat acatgactcc atggctgtca aatctagttc gtttttgcaa
     6781 gcataatgat gatgccaaac gacaaagagt tactgctatt cttgacaagc tgataacaat
     6841 gaccatcaat gaaaaacaga tgtacccatc tattcaagca aaaatatggg gaagccttgg
     6901 gcagattaca gatctgcttg atgttgtact agacagtttc atcaaaacca gtgcaacagg
     6961 tggcttggga tcaataaaag ctgaggtgat ggcagatact gctgtagctt tggcttctgg
     7021 aaatgtgaaa ttggtttcaa gcaaggttat tggaaggatg tgcaaaataa ttgacaagac
     7081 atgcttatct ccaactccta ctttagaaca acatcttatg tgggatgata ttgctatttt
     7141 agcacgctac atgctgatgc tgtccttcaa caattccctt gatgtggcag ctcatcttcc
     7201 ctacctcttc cacgttgtta ctttcttagt agccacaggt ccgctctccc ttagagcttc
     7261 cacacatgga ctggtcatta atatcattca ctctctgtgt acttgttcac agcttcattt
     7321 tagtgaagag accaagcaag ttttgagact cagtctgaca gagttctcat tacccaaatt
     7381 ttacttgctg tttggcatta gcaaagtcaa gtcagctgct gtcattgcct tccgttccag
     7441 ttaccgggac aggtcattct ctcctggctc ctatgagaga gagacttttg ctttgacatc
     7501 cttggaaaca gtcacagaag ctttgttgga gatcatggag gcatgcatga gagatattcc
     7561 aacgtgcaag tggctggacc agtggacaga actagctcaa agatttgcat tccaatataa
     7621 tccatccctg caaccaagag ctcttgttgt ctttgggtgt attagcaaac gagtgtctca
     7681 tgggcagata aagcagataa tccgtattct tagcaaggca cttgagagtt gcttaaaagg
     7741 acctgacact tacaacagtc aagttctgat agaagctaca gtaatagcac taaccaaatt
     7801 acagccactt cttaataagg actcgcctct gcacaaagcc ctcttttggg tagctgtggc
     7861 tgtgctgcag cttgatgagg tcaacttgta ttcagcaggt accgcacttc ttgaacaaaa
     7921 cctgcatact ttagatagtc tccgtatatt caatgacaag agtccagagg aagtatttat
     7981 ggcaatccgg aatcctctgg agtggcactg caagcaaatg gatcattttg ttggactcaa
     8041 tttcaactct aactttaact ttgcattggt tggacacctt ttaaaagggt acaggcatcc
     8101 ttcacctgct attgttgcaa gaacagtcag aattttacat acactactaa ctctggttaa
     8161 caaacacaga aattgtgaca aatttgaagt gaatacacag agcgtggcct acttagcagc
     8221 tttacttaca gtgtctgaag aagttcgaag tcgctgcagc ctaaaacata gaaagtcact
     8281 tcttcttact gatatttcaa tggaaaatgt tcctatggat acatatccca ttcatcatgg
     8341 tgacccttcc tataggacac taaaggagac tcagccatgg tcctctccca aaggttctga
     8401 aggatacctt gcagccacct atccaactgt cggccagacc agtccccgag ccaggaaatc
     8461 catgagcctg gacatggggc aaccttctca ggccaacact aagaagttgc ttggaacaag
     8521 gaaaagtttt gatcacttga tatcagacac aaaggctcct aaaaggcaag aaatggaatc
     8581 agggatcaca acacccccca aaatgaggag agtagcagaa actgattatg aaatggaaac
     8641 tcagaggatt tcctcatcac aacagcaccc acatttacgt aaagtttcag tgtctgaatc
     8701 aaatgttctc ttggatgaag aagtacttac tgatccgaag atccaggcgc tgcttcttac
     8761 tgttctagct acactggtaa aatataccac agatgagttt gatcaacgaa ttctttatga
     8821 atacttagca gaggccagtg ttgtgtttcc caaagtcttt cctgttgtgc ataatttgtt
     8881 ggactctaag atcaacaccc tgttatcatt gtgccaagat ccaaatttgt taaatccaat
     8941 ccatggaatt gtgcagagtg tggtgtacca tgaagaatcc ccaccacaat accaaacatc
     9001 ttacctgcaa agttttggtt ttaatggctt gtggcggttt gcaggaccgt tttcaaagca
     9061 aacacaaatt ccagactatg ctgagcttat tgttaagttt cttgatgcct tgattgacac
     9121 gtacctgcct ggaattgatg aagaaaccag tgaagaatcc ctcctgactc ccacatctcc
     9181 ttaccctcct gcactgcaga gccagcttag tatcactgcc aaccttaacc tttctaattc
     9241 catgacctca cttgcaactt cccagcattc cccaggaatc gacaaggaga acgttgaact
     9301 ctcccctacc actggccact gtaacagtgg acgaactcgc cacggatccg caagccaagt
     9361 gcagaagcaa agaagcgcag gatcatttaa gagaaatagc attaagaaga tcgtgtagcg
     9421 gccgcatcgt gactgactga cgatctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     9481 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     9541 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     9601 gtcacgtagc gatagcggag tgtataattc ttgaagacga aagggcctcg tgatacgcct
     9661 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     9721 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     9781 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     9841 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     9901 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     9961 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
    10021 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
    10081 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
    10141 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
    10201 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
    10261 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
    10321 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
    10381 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
    10441 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
    10501 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
    10561 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
    10621 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
    10681 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
    10741 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataa
//
