LOCUS       sequence-229898-       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)
FEATURES             Location/Qualifiers
     source          1..10788
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      72..660
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     561..580
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(9471..9493)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9638..9656)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9724..9828
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9829..9897,9898..10689)
                     /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(10047..10066)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
       61 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
      121 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
      181 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
      241 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
      301 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
      361 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
      421 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
      481 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
      541 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
      601 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
      661 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
      721 tcgtacgacg cgtcccggga ccggtggcat ggccgcgcac aggccggtgg aatgggtcca
      781 ggccgtggtc agccgcttcg acgagcagct tccaataaaa acaggacagc agaacacaca
      841 taccaaagtc agtactgagc acaacaagga atgtctaatc aatatttcca aatacaagtt
      901 ttctttggtt ataagcggcc tcactactat tttaaagaat gttaacaata tgagaatatt
      961 tggagaagct gctgaaaaaa atttatatct ctctcagttg attatattgg atacactgga
     1021 aaaatgtctt gctgggcaac caaaggacac aatgagatta gatgaaacga tgctggtcaa
     1081 acagttgctg ccagaaatct gccattttct tcacacctgt cgtgaaggaa accagcatgc
     1141 agctgaactt cggaattctg cctctggggt tttattttct ctcagctgca acaacttcaa
     1201 tgcagtcttt agtcgcattt ctaccaggtt acaggaatta actgtttgtt cagaagacaa
     1261 tgttgatgtt catgatatag aattgttaca gtatatcaat gtggattgtg caaaattaaa
     1321 acgactcctg aaggaaacag catttaaatt taaagcccta aagaaggttg cgcagttagc
     1381 agttataaat agcctggaaa aggcattttg gaactgggta gaaaattatc cagatgaatt
     1441 tacaaaacta taccagatcc cacagactga tatggctgaa tgtgcagaaa agctatttga
     1501 cttggtggat ggttttgctg aaagcaccaa acgtaaagca gcagtttggc cactacaaat
     1561 cattctcctt atcttgtgtc cagaaataat ccaggatata tccaaagacg tggttgatga
     1621 aaacaacatg aataagaagt tatttctgga cagtctacga aaagctcttg ctggccatgg
     1681 aggaagtagg cagctgacag aaagtgctgc aattgcctgt gtcaaactgt gtaaagcaag
     1741 tacttacatc aattgggaag ataactctgt cattttccta cttgttcagt ccatggtggt
     1801 tgatcttaag aacctgcttt ttaatccaag taagccattc tcaagaggca gtcagcctgc
     1861 agatgtggat ctaatgattg actgccttgt ttcttgcttt cgtataagcc ctcacaacaa
     1921 ccaacacttt aagatctgcc tggctcagaa ttcaccttct acatttcact atgtgctggt
     1981 aaattcactc catcgaatca tcaccaattc cgcattggat tggtggccta agattgatgc
     2041 tgtgtattgt cactcggttg aacttcgaaa tatgtttggt gaaacacttc ataaagcagt
     2101 gcaaggttgt ggagcacacc cagcaatacg aatggcaccg agtcttacat ttaaagaaaa
     2161 agtaacaagc cttaaattta aagaaaaacc tacagacctg gagacaagaa gctataagta
     2221 tcttctcttg tccatggtga aactaattca tgcagatcca aagctcttgc tttgtaatcc
     2281 aagaaaacag gggcccgaaa cccaaggcag tacagcagaa ttaattacag ggctcgtcca
     2341 actggtccct cagtcacaca tgccagagat tgctcaggaa gcaatggagg ctctgctggt
     2401 tcttcatcag ttagatagca ttgatttgtg gaatcctgat gctcctgtag aaacattttg
     2461 ggagattagc tcacaaatgc ttttttacat ctgcaagaaa ttaactagtc atcaaatgct
     2521 tagtagcaca gaaattctca agtggttgcg ggaaatattg atctgcagga ataaatttct
     2581 tcttaaaaat aagcaggcag atagaagttc ctgtcacttt ctcctttttt acggggtagg
     2641 atgtgatatt ccttctagtg gaaataccag tcaaatgtcc atggatcatg aagaattact
     2701 acgtactcct ggagcctctc tccggaaggg aaaagggaac tcctctatgg atagtgcagc
     2761 aggatgcagc ggaacccccc caatttgccg acaagcccag accaaactag aagtggccct
     2821 gtacatgttt ctgtggaacc ctgacactga agctgttctg gttgccatgt cctgtttccg
     2881 ccacctctgt gaggaagcag atatccggtg tggggtggat gaagtgtcag tgcataacct
     2941 cttgcccaac tataacacat tcatggagtt tgcctctgtc agcaatatga tgtcaacagg
     3001 aagagcagca cttcagaaaa gagtgatggc actgctgagg cgcattgagc atcccactgc
     3061 aggaaacact gaggcttggg aagatacaca tgcaaaatgg gaacaagcaa caaagctaat
     3121 ccttaactat ccaaaagcca aaatggaaga tggccaggct gctgaaagcc ttcacaagac
     3181 cattgtcaag aggcgaatgt cccatgtgag tggaggagga tccatagatt tgtctgacac
     3241 agactcccta caggaatgga tcaacatgac tggcttcctt tgtgcccttg ggggagtgtg
     3301 cctccagcag agaagcaatt ctggcctggc aacctatagc ccacccatgg gtccagtcag
     3361 tgaacgtaag ggttctatga tttcagtgat gtcttcagag ggaaacgcag atacacctgt
     3421 cagcaaattt atggatcggc tgttgtcctt aatggtgtgt aaccatgaga aagtgggact
     3481 tcaaatacgg accaatgtta aggatctggt gggtctagaa ttgagtcctg ctctgtatcc
     3541 aatgctattt aacaaattga agaataccat cagcaagttt tttgactccc aaggacaggt
     3601 tttattgact gataccaata ctcaatttgt agaacaaacc atagctataa tgaagaactt
     3661 gctagataat catactgaag gcagctctga acatctaggg caagctagca ttgaaacaat
     3721 gatgttaaat ctggtcaggt atgttcgtgt gcttgggaat atggtccatg caattcaaat
     3781 aaaaacgaaa ctgtgtcaat tagttgaagt aatgatggca aggagagatg acctctcatt
     3841 ttgccaagag atgaaattta ggaataagat ggtagaatac ctgacagact gggttatggg
     3901 aacatcaaac caagcagcag atgatgatgt aaaatgtctt acaagagatt tggaccaggc
     3961 aagcatggaa gcagtagttt cacttctagc tggtctccct ctgcagcctg aagaaggaga
     4021 tggtgtggaa ttgatggaag ccaaatcaca gttatttctt aaatacttca cattatttat
     4081 gaaccttttg aatgactgca gtgaagttga agatgaaagt gcgcaaacag gtggcaggaa
     4141 acgtggcatg tctcggaggc tggcatcact gaggcactgt acggtccttg caatgtcaaa
     4201 cttactcaat gccaacgtag acagtggtct catgcactcc ataggcttag gttaccacaa
     4261 ggatctccag acaagagcta catttatgga agttctgaca aaaatccttc aacaaggcac
     4321 agaatttgac acacttgcag aaacagtatt ggctgatcgg tttgagagat tggtggaact
     4381 ggtcacaatg atgggtgatc aaggagaact ccctatagcg atggctctgg ccaatgtggt
     4441 tccttgttct cagtgggatg aactagctcg agttctggtt actctgtttg attctcggca
     4501 tttactctac caactgctct ggaacatgtt ttctaaagaa gtagaattgg cagactccat
     4561 gcagactctc ttccgaggca acagcttggc cagtaaaata atgacattct gtttcaaggt
     4621 atatggtgct acctatctac aaaaactcct ggatccttta ttacgaattg tgatcacatc
     4681 ctctgattgg caacatgtta gctttgaagt ggatcctacc aggttagaac catcagagag
     4741 ccttgaggaa aaccagcgga acctccttca gatgactgaa aagttcttcc atgccatcat
     4801 cagttcctcc tcagaattcc cccctcaact tcgaagtgtg tgccactgtt tataccaggc
     4861 aacttgccac tccctactga ataaagctac agtaaaagaa aaaaaggaaa acaaaaaatc
     4921 agtggttagc cagcgtttcc ctcagaacag catcggtgca gtaggaagtg ccatgttcct
     4981 cagatttatc aatcctgcca ttgtctcacc gtatgaagca gggattttag ataaaaagcc
     5041 accacctaga atcgaaaggg gcttgaagtt aatgtcaaag atacttcaga gtattgccaa
     5101 tcatgttctc ttcacaaaag aagaacatat gcggcctttc aatgattttg tgaaaagcaa
     5161 ctttgatgca gcacgcaggt ttttccttga tatagcatct gattgtccta caagtgatgc
     5221 agtaaatcat agtctttcct tcataagtga cggcaatgtg cttgctttac atcgtctact
     5281 ctggaacaat caggagaaaa ttgggcagta tctttccagc aacagggatc ataaagctgt
     5341 tggaagacga ccttttgata agatggcaac acttcttgca tacctgggtc ctccagagca
     5401 caaacctgtg gcagatacac actggtccag ccttaacctt accagttcaa agtttgagga
     5461 atttatgact aggcatcagg tacatgaaaa agaagaattc aaggctttga aaacgttaag
     5521 tattttctac caagctggga cttccaaagc tgggaatcct attttttatt atgttgcacg
     5581 gaggtaagaa atactatgtt ttgggtctct taacagaatt ttttaaatta tagcaaatat
     5641 agagagatgg caactagatt agtggcatct gtatatttat tttaaacact gctaataatc
     5701 tttgtctttt ttgtcatttt ccttaggttc aaaactggtc aaatcaatgg tgatttgctg
     5761 atataccatg tcttactgac tttaaagcca tattatgcaa agccatatga aattgtagtg
     5821 gaccttaccc ataccgggcc tagcaatcgc tttaaaacag actttctctc taagtggttt
     5881 gttgtttttc ctggctttgc ttacgacaac gtctccgcag tctatatcta taactgtaac
     5941 tcctgggtca gggagtacac caagtatcat gagcggctgc tgactggcct caaaggtagc
     6001 aaaaggcttg ttttcataga ctgtcctggg aaactggctg agcacataga gcatgaacaa
     6061 cagaaactac ctgctgccac cttggcttta gaagaggacc tgaaggtatt ccacaatgct
     6121 ctcaagctag ctcacaaaga caccaaagtt tctattaaag ttggttctac tgctgtccaa
     6181 gtaacttcag cagagcgaac aaaagtccta gggcaatcag tctttctaaa tgacatttat
     6241 tatgcttcgg aaattgaaga aatctgccta gtagatgaga accagttcac cttaaccatt
     6301 gcaaaccagg gcacgccgct caccttcatg caccaggagt gtgaagccat tgtccagtct
     6361 atcattcata tccggacccg ctgggaactg tcacagcccg actctatccc ccaacacacc
     6421 aagattcggc caaaagatgt ccctgggaca ctgctcaata tcgcattact taatttaggc
     6481 agttctgacc cgagtttacg gtcagctgcc tataatcttc tgtgtgcctt aacttgtacc
     6541 tttaatttaa aaatcgaggg ccagttacta gagacatcag gtttatgtat ccctgccaac
     6601 aacaccctct ttattgtctc tattagtaag acactggcag ccaatgagcc acacctcacg
     6661 ttagaatttt tggaagagtg tatttctgga tttagcaaat ctagtattga attgaaacac
     6721 ctttgtttgg aatacatgac tccatggctg tcaaatctag ttcgtttttg caagcataat
     6781 gatgatgcca aacgacaaag agttactgct attcttgaca agctgataac aatgaccatc
     6841 aatgaaaaac agatgtaccc atctattcaa gcaaaaatat ggggaagcct tgggcagatt
     6901 acagatctgc ttgatgttgt actagacagt ttcatcaaaa ccagtgcaac aggtggcttg
     6961 ggatcaataa aagctgaggt gatggcagat actgctgtag ctttggcttc tggaaatgtg
     7021 aaattggttt caagcaaggt tattggaagg atgtgcaaaa taattgacaa gacatgctta
     7081 tctccaactc ctactttaga acaacatctt atgtgggatg atattgctat tttagcacgc
     7141 tacatgctga tgctgtcctt caacaattcc cttgatgtgg cagctcatct tccctacctc
     7201 ttccacgttg ttactttctt agtagccaca ggtccgctct cccttagagc ttccacacat
     7261 ggactggtca ttaatatcat tcactctctg tgtacttgtt cacagcttca ttttagtgaa
     7321 gagaccaagc aagttttgag actcagtctg acagagttct cattacccaa attttacttg
     7381 ctgtttggca ttagcaaagt caagtcagct gctgtcattg ccttccgttc cagttaccgg
     7441 gacaggtcat tctctcctgg ctcctatgag agagagactt ttgctttgac atccttggaa
     7501 acagtcacag aagctttgtt ggagatcatg gaggcatgca tgagagatat tccaacgtgc
     7561 aagtggctgg accagtggac agaactagct caaagatttg cattccaata taatccatcc
     7621 ctgcaaccaa gagctcttgt tgtctttggg tgtattagca aacgagtgtc tcatgggcag
     7681 ataaagcaga taatccgtat tcttagcaag gcacttgaga gttgcttaaa aggacctgac
     7741 acttacaaca gtcaagttct gatagaagct acagtaatag cactaaccaa attacagcca
     7801 cttcttaata aggactcgcc tctgcacaaa gccctctttt gggtagctgt ggctgtgctg
     7861 cagcttgatg aggtcaactt gtattcagca ggtaccgcac ttcttgaaca aaacctgcat
     7921 actttagata gtctccgtat attcaatgac aagagtccag aggaagtatt tatggcaatc
     7981 cggaatcctc tggagtggca ctgcaagcaa atggatcatt ttgttggact caatttcaac
     8041 tctaacttta actttgcatt ggttggacac cttttaaaag ggtacaggca tccttcacct
     8101 gctattgttg caagaacagt cagaatttta catacactac taactctggt taacaaacac
     8161 agaaattgtg acaaatttga agtgaataca cagagcgtgg cctacttagc agctttactt
     8221 acagtgtctg aagaagttcg aagtcgctgc agcctaaaac atagaaagtc acttcttctt
     8281 actgatattt caatggaaaa tgttcctatg gatacatatc ccattcatca tggtgaccct
     8341 tcctatagga cactaaagga gactcagcca tggtcctctc ccaaaggttc tgaaggatac
     8401 cttgcagcca cctatccaac tgtcggccag accagtcccc gagccaggaa atccatgagc
     8461 ctggacatgg ggcaaccttc tcaggccaac actaagaagt tgcttggaac aaggaaaagt
     8521 tttgatcact tgatatcaga cacaaaggct cctaaaaggc aagaaatgga atcagggatc
     8581 acaacacccc ccaaaatgag gagagtagca gaaactgatt atgaaatgga aactcagagg
     8641 atttcctcat cacaacagca cccacattta cgtaaagttt cagtgtctga atcaaatgtt
     8701 ctcttggatg aagaagtact tactgatccg aagatccagg cgctgcttct tactgttcta
     8761 gctacactgg taaaatatac cacagatgag tttgatcaac gaattcttta tgaatactta
     8821 gcagaggcca gtgttgtgtt tcccaaagtc tttcctgttg tgcataattt gttggactct
     8881 aagatcaaca ccctgttatc attgtgccaa gatccaaatt tgttaaatcc aatccatgga
     8941 attgtgcaga gtgtggtgta ccatgaagaa tccccaccac aataccaaac atcttacctg
     9001 caaagttttg gttttaatgg cttgtggcgg tttgcaggac cgttttcaaa gcaaacacaa
     9061 attccagact atgctgagct tattgttaag tttcttgatg ccttgattga cacgtacctg
     9121 cctggaattg atgaagaaac cagtgaagaa tccctcctga ctcccacatc tccttaccct
     9181 cctgcactgc agagccagct tagtatcact gccaacctta acctttctaa ttccatgacc
     9241 tcacttgcaa cttcccagca ttccccagga atcgacaagg agaacgttga actctcccct
     9301 accactggcc actgtaacag tggacgaact cgccacggat ccgcaagcca agtgcagaag
     9361 caaagaagcg ctggcagttt caaacgtaat agcattaaga agatcgtgta gcggccgcat
     9421 cgtgactgac tgacgatctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     9481 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     9541 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
     9601 agcgatagcg gagtgtataa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     9661 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     9721 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     9781 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     9841 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     9901 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     9961 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
    10021 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
    10081 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
    10141 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
    10201 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
    10261 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
    10321 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
    10381 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
    10441 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
    10501 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
    10561 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
    10621 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
    10681 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
    10741 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatg
//
