LOCUS       sequence-229906-       10725 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 10725)
  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 10725)
  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 T1 (+minintr)
FEATURES             Location/Qualifiers
     source          1..10725
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(8711..8733)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8878..8896)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8964..9068
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9069..9137,9138..9929)
                     /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(9287..9306)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10100..10688
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10589..10608
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttgctggcc ttttgctcac atgtcgtacg acgcgtcccg ggaccggtgg catggccgcg
       61 cacaggccgg tggaatgggt ccaggccgtg gtcagccgct tcgacgagca gcttccaata
      121 aaaacaggac agcagaacac acataccaaa gtcagtactg agcacaacaa ggaatgtcta
      181 atcaatattt ccaaatacaa gttttctttg gttataagcg gcctcactac tattttaaag
      241 aatgttaaca atatgagaat atttggagaa gctgctgaaa aaaatttata tctctctcag
      301 ttgattatat tggatacact ggaaaaatgt cttgctgggc aaccaaagga cacaatgaga
      361 ttagatgaaa cgatgctggt caaacagttg ctgccagaaa tctgccattt tcttcacacc
      421 tgtcgtgaag gaaaccagca tgcagctgaa cttcggaatt ctgcctctgg ggttttattt
      481 tctctcagct gcaacaactt caatgcagtc tttagtcgca tttctaccag gttacaggaa
      541 ttaactgttt gttcagaaga caatgttgat gttcatgata tagaattgtt acagtatatc
      601 aatgtggatt gtgcaaaatt aaaacgactc ctgaaggaaa cagcatttaa atttaaagcc
      661 ctaaagaagg ttgcgcagtt agcagttata aatagcctgg aaaaggcatt ttggaactgg
      721 gtagaaaatt atccagatga atttacaaaa ctataccaga tcccacagac tgatatggct
      781 gaatgtgcag aaaagctatt tgacttggtg gatggttttg ctgaaagcac caaacgtaaa
      841 gcagcagttt ggccactaca aatcattctc cttatcttgt gtccagaaat aatccaggat
      901 atatccaaag acgtggttga tgaaaacaac atgaataaga agttatttct ggacagtcta
      961 cgaaaagctc ttgctggcca tggaggaagt aggcagctga cagaaagtgc tgcaattgcc
     1021 tgtgtcaaac tgtgtaaagc aagtacttac atcaattggg aagataactc tgtcattttc
     1081 ctacttgttc agtccatggt ggttgatctt aagaacctgc tttttaatcc aagtaagcca
     1141 ttctcaagag gcagtcagcc tgcagatgtg gatctaatga ttgactgcct tgtttcttgc
     1201 tttcgtataa gccctcacaa caaccaacac tttaagatct gcctggctca gaattcacct
     1261 tctacatttc actatgtgct ggtaaattca ctccatcgaa tcatcaccaa ttccgcattg
     1321 gattggtggc ctaagattga tgctgtgtat tgtcactcgg ttgaacttcg aaatatgttt
     1381 ggtgaaacac ttcataaagc agtgcaaggt tgtggagcac acccagcaat acgaatggca
     1441 ccgagtctta catttaaaga aaaagtaaca agccttaaat ttaaagaaaa acctacagac
     1501 ctggagacaa gaagctataa gtatcttctc ttgtccatgg tgaaactaat tcatgcagat
     1561 ccaaagctct tgctttgtaa tccaagaaaa caggggcccg aaacccaagg cagtacagca
     1621 gaattaatta cagggctcgt ccaactggtc cctcagtcac acatgccaga gattgctcag
     1681 gaagcaatgg aggctctgct ggttcttcat cagttagata gcattgattt gtggaatcct
     1741 gatgctcctg tagaaacatt ttgggagatt agctcacaaa tgctttttta catctgcaag
     1801 aaattaacta gtcatcaaat gcttagtagc acagaaattc tcaagtggtt gcgggaaata
     1861 ttgatctgca ggaataaatt tcttcttaaa aataagcagg cagatagaag ttcctgtcac
     1921 tttctccttt tttacggggt aggatgtgat attccttcta gtggaaatac cagtcaaatg
     1981 tccatggatc atgaagaatt actacgtact cctggagcct ctctccggaa gggaaaaggg
     2041 aactcctcta tggatagtgc agcaggatgc agcggaaccc ccccaatttg ccgacaagcc
     2101 cagaccaaac tagaagtggc cctgtacatg tttctgtgga accctgacac tgaagctgtt
     2161 ctggttgcca tgtcctgttt ccgccacctc tgtgaggaag cagatatccg gtgtggggtg
     2221 gatgaagtgt cagtgcataa cctcttgccc aactataaca cattcatgga gtttgcctct
     2281 gtcagcaata tgatgtcaac aggaagagca gcacttcaga aaagagtgat ggcactgctg
     2341 aggcgcattg agcatcccac tgcaggaaac actgaggctt gggaagatac acatgcaaaa
     2401 tgggaacaag caacaaagct aatccttaac tatccaaaag ccaaaatgga agatggccag
     2461 gctgctgaaa gccttcacaa gaccattgtc aagaggcgaa tgtcccatgt gagtggagga
     2521 ggatccatag atttgtctga cacagactcc ctacaggaat ggatcaacat gactggcttc
     2581 ctttgtgccc ttgggggagt gtgcctccag cagagaagca attctggcct ggcaacctat
     2641 agcccaccca tgggtccagt cagtgaacgt aagggttcta tgatttcagt gatgtcttca
     2701 gagggaaacg cagatacacc tgtcagcaaa tttatggatc ggctgttgtc cttaatggtg
     2761 tgtaaccatg agaaagtggg acttcaaata cggaccaatg ttaaggatct ggtgggtcta
     2821 gaattgagtc ctgctctgta tccaatgcta tttaacaaat tgaagaatac catcagcaag
     2881 ttttttgact cccaaggaca ggttttattg actgatacca atactcaatt tgtagaacaa
     2941 accatagcta taatgaagaa cttgctagat aatcatactg aaggcagctc tgaacatcta
     3001 gggcaagcta gcattgaaac aatgatgtta aatctggtca ggtatgttcg tgtgcttggg
     3061 aatatggtcc atgcaattca aataaaaacg aaactgtgtc aattagttga agtaatgatg
     3121 gcaaggagag atgacctctc attttgccaa gagatgaaat ttaggaataa gatggtagaa
     3181 tacctgacag actgggttat gggaacatca aaccaagcag cagatgatga tgtaaaatgt
     3241 cttacaagag atttggacca ggcaagcatg gaagcagtag tttcacttct agctggtctc
     3301 cctctgcagc ctgaagaagg agatggtgtg gaattgatgg aagccaaatc acagttattt
     3361 cttaaatact tcacattatt tatgaacctt ttgaatgact gcagtgaagt tgaagatgaa
     3421 agtgcgcaaa caggtggcag gaaacgtggc atgtctcgga ggctggcatc actgaggcac
     3481 tgtacggtcc ttgcaatgtc aaacttactc aatgccaacg tagacagtgg tctcatgcac
     3541 tccataggct taggttacca caaggatctc cagacaagag ctacatttat ggaagttctg
     3601 acaaaaatcc ttcaacaagg cacagaattt gacacacttg cagaaacagt attggctgat
     3661 cggtttgaga gattggtgga actggtcaca atgatgggtg atcaaggaga actccctata
     3721 gcgatggctc tggccaatgt ggttccttgt tctcagtggg atgaactagc tcgagttctg
     3781 gttactctgt ttgattctcg gcatttactc taccaactgc tctggaacat gttttctaaa
     3841 gaagtagaat tggcagactc catgcagact ctcttccgag gcaacagctt ggccagtaaa
     3901 ataatgacat tctgtttcaa ggtatatggt gctacctatc tacaaaaact cctggatcct
     3961 ttattacgaa ttgtgatcac atcctctgat tggcaacatg ttagctttga agtggatcct
     4021 accaggttag aaccatcaga gagccttgag gaaaaccagc ggaacctcct tcagatgact
     4081 gaaaagttct tccatgccat catcagttcc tcctcagaat tcccccctca acttcgaagt
     4141 gtgtgccact gtttatacca ggtggttagc cagcgtttcc ctcagaacag catcggtgca
     4201 gtaggaagtg ccatgttcct cagatttatc aatcctgcca ttgtctcacc gtatgaagca
     4261 gggattttag ataaaaagcc accacctaga atcgaaaggg gcttgaagtt aatgtcaaag
     4321 atacttcaga gtattgccaa tcatgttctc ttcacaaaag aagaacatat gcggcctttc
     4381 aatgattttg tgaaaagcaa ctttgatgca gcacgcaggt ttttccttga tatagcatct
     4441 gattgtccta caagtgatgc agtaaatcat agtctttcct tcataagtga cggcaatgtg
     4501 cttgctttac atcgtctact ctggaacaat caggagaaaa ttgggcagta tctttccagc
     4561 aacagggatc ataaagctgt tggaagacga ccttttgata agatggcaac acttcttgca
     4621 tacctgggtc ctccagagca caaacctgtg gcagatacac actggtccag ccttaacctt
     4681 accagttcaa agtttgagga atttatgact aggcatcagg tacatgaaaa agaagaattc
     4741 aaggctttga aaacgttaag tattttctac caagctggga cttccaaagc tgggaatcct
     4801 attttttatt atgttgcacg gaggtaagaa atactatgtt ttgggtctct taacagaatt
     4861 ttttaaatta tagcaaatat agagagatgg caactagatt agtggcatct gtatatttat
     4921 tttaaacact gctaataatc tttgtctttt ttgtcatttt ccttaggttc aaaactggtc
     4981 aaatcaatgg tgatttgctg atataccatg tcttactgac tttaaagcca tattatgcaa
     5041 agccatatga aattgtagtg gaccttaccc ataccgggcc tagcaatcgc tttaaaacag
     5101 actttctctc taagtggttt gttgtttttc ctggctttgc ttacgacaac gtctccgcag
     5161 tctatatcta taactgtaac tcctgggtca gggagtacac caagtatcat gagcggctgc
     5221 tgactggcct caaaggtagc aaaaggcttg ttttcataga ctgtcctggg aaactggctg
     5281 agcacataga gcatgaacaa cagaaactac ctgctgccac cttggcttta gaagaggacc
     5341 tgaaggtatt ccacaatgct ctcaagctag ctcacaaaga caccaaagtt tctattaaag
     5401 ttggttctac tgctgtccaa gtaacttcag cagagcgaac aaaagtccta gggcaatcag
     5461 tctttctaaa tgacatttat tatgcttcgg aaattgaaga aatctgccta gtagatgaga
     5521 accagttcac cttaaccatt gcaaaccagg gcacgccgct caccttcatg caccaggagt
     5581 gtgaagccat tgtccagtct atcattcata tccggacccg ctgggaactg tcacagcccg
     5641 actctatccc ccaacacacc aagattcggc caaaagatgt ccctgggaca ctgctcaata
     5701 tcgcattact taatttaggc agttctgacc cgagtttacg gtcagctgcc tataatcttc
     5761 tgtgtgcctt aacttgtacc tttaatttaa aaatcgaggg ccagttacta gagacatcag
     5821 gtttatgtat ccctgccaac aacaccctct ttattgtctc tattagtaag acactggcag
     5881 ccaatgagcc acacctcacg ttagaatttt tggaagagtg tatttctgga tttagcaaat
     5941 ctagtattga attgaaacac ctttgtttgg aatacatgac tccatggctg tcaaatctag
     6001 ttcgtttttg caagcataat gatgatgcca aacgacaaag agttactgct attcttgaca
     6061 agctgataac aatgaccatc aatgaaaaac agatgtaccc atctattcaa gcaaaaatat
     6121 ggggaagcct tgggcagatt acagatctgc ttgatgttgt actagacagt ttcatcaaaa
     6181 ccagtgcaac aggtggcttg ggatcaataa aagctgaggt gatggcagat actgctgtag
     6241 ctttggcttc tggaaatgtg aaattggttt caagcaaggt tattggaagg atgtgcaaaa
     6301 taattgacaa gacatgctta tctccaactc ctactttaga acaacatctt atgtgggatg
     6361 atattgctat tttagcacgc tacatgctga tgctgtcctt caacaattcc cttgatgtgg
     6421 cagctcatct tccctacctc ttccacgttg ttactttctt agtagccaca ggtccgctct
     6481 cccttagagc ttccacacat ggactggtca ttaatatcat tcactctctg tgtacttgtt
     6541 cacagcttca ttttagtgaa gagaccaagc aagttttgag actcagtctg acagagttct
     6601 cattacccaa attttacttg ctgtttggca ttagcaaagt caagtcagct gctgtcattg
     6661 ccttccgttc cagttaccgg gacaggtcat tctctcctgg ctcctatgag agagagactt
     6721 ttgctttgac atccttggaa acagtcacag aagctttgtt ggagatcatg gaggcatgca
     6781 tgagagatat tccaacgtgc aagtggctgg accagtggac agaactagct caaagatttg
     6841 cattccaata taatccatcc ctgcaaccaa gagctcttgt tgtctttggg tgtattagca
     6901 aacgagtgtc tcatgggcag ataaagcaga taatccgtat tcttagcaag gcacttgaga
     6961 gttgcttaaa aggacctgac acttacaaca gtcaagttct gatagaagct acagtaatag
     7021 cactaaccaa attacagcca cttcttaata aggactcgcc tctgcacaaa gccctctttt
     7081 gggtagctgt ggctgtgctg cagcttgatg aggtcaactt gtattcagca ggtaccgcac
     7141 ttcttgaaca aaacctgcat actttagata gtctccgtat attcaatgac aagagtccag
     7201 aggaagtatt tatggcaatc cggaatcctc tggagtggca ctgcaagcaa atggatcatt
     7261 ttgttggact caatttcaac tctaacttta actttgcatt ggttggacac cttttaaaag
     7321 ggtacaggca tccttcacct gctattgttg caagaacagt cagaatttta catacactac
     7381 taactctggt taacaaacac agaaattgtg acaaatttga agtgaataca cagagcgtgg
     7441 cctacttagc agctttactt acagtgtctg aagaagttcg aagtcgctgc agcctaaaac
     7501 atagaaagtc acttcttctt actgatattt caatggaaaa tgttcctatg gatacatatc
     7561 ccattcatca tggtgaccct tcctatagga cactaaagga gactcagcca tggtcctctc
     7621 ccaaaggttc tgaaggatac cttgcagcca cctatccaac tgtcggccag accagtcccc
     7681 gagccaggaa atccatgagc ctggacatgg ggcaaccttc tcaggccaac actaagaagt
     7741 tgcttggaac aaggaaaagt tttgatcact tgatatcaga cacaaaggct cctaaaaggc
     7801 aagaaatgga atcagggatc acaacacccc ccaaaatgag gagagtagca gaaactgatt
     7861 atgaaatgga aactcagagg atttcctcat cacaacagca cccacattta cgtaaagttt
     7921 cagtgtctga atcaaatgtt ctcttggatg aagaagtact tactgatccg aagatccagg
     7981 cgctgcttct tactgttcta gctacactgg taaaatatac cacagatgag tttgatcaac
     8041 gaattcttta tgaatactta gcagaggcca gtgttgtgtt tcccaaagtc tttcctgttg
     8101 tgcataattt gttggactct aagatcaaca ccctgttatc attgtgccaa gatccaaatt
     8161 tgttaaatcc aatccatgga attgtgcaga gtgtggtgta ccatgaagaa tccccaccac
     8221 aataccaaac atcttacctg caaagttttg gttttaatgg cttgtggcgg tttgcaggac
     8281 cgttttcaaa gcaaacacaa attccagact atgctgagct tattgttaag tttcttgatg
     8341 ccttgattga cacgtacctg cctggaattg atgaagaaac cagtgaagaa tccctcctga
     8401 ctcccacatc tccttaccct cctgcactgc agagccagct tagtatcact gccaacctta
     8461 acctttctaa ttccatgacc tcacttgcaa cttcccagca ttccccagga atcgacaagg
     8521 agaacgttga actctcccct accactggcc actgtaacag tggacgaact cgccacggat
     8581 ccgcaagcca agtgcagaag caaagaagcg ctggcagttt caaacgtaat agcattaaga
     8641 agatcgtgta gcggccgcat cgtgactgac tgacgatctg cctcgcgcgt ttcggtgatg
     8701 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     8761 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg
     8821 cagccatgac ccagtcacgt agcgatagcg gagtgtataa ttcttgaaga cgaaagggcc
     8881 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     8941 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     9001 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     9061 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     9121 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     9181 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     9241 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     9301 tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     9361 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     9421 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     9481 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     9541 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     9601 ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     9661 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     9721 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     9781 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     9841 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     9901 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     9961 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
    10021 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
    10081 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
    10141 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
    10201 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
    10261 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
    10321 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
    10381 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
    10441 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
    10501 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
    10561 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
    10621 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
    10681 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggcct
//
