LOCUS       sequence-225452-       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"
     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(9415..9437)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9582..9600)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9668..9772
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9773..9841,9842..10633)
                     /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(9991..10010)
                     /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 cgaggcaaca 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 accaggtggt tagccagcgt ttccctcaga acagcatcgg tgcagtagga agtgccatgt
     4921 tcctcagatt tatcaatcct gccattgtct caccgtatga agcagggatt ttagataaaa
     4981 agccaccacc tagaatcgaa aggggcttga agttaatgtc aaagatactt cagagtattg
     5041 ccaatcatgt tctcttcaca aaagaagaac atatgcggcc tttcaatgat tttgtgaaaa
     5101 gcaactttga tgcagcacgc aggtttttcc ttgatatagc atctgattgt cctacaagtg
     5161 atgcagtaaa tcatagtctt tccttcataa gtgacggcaa tgtgcttgct ttacatcgtc
     5221 tactctggaa caatcaggag aaaattgggc agtatctttc cagcaacagg gatcataaag
     5281 ctgttggaag acgacctttt gataagatgg caacacttct tgcatacctg ggtcctccag
     5341 agcacaaacc tgtggcagat acacactggt ccagccttaa ccttaccagt tcaaagtttg
     5401 aggaatttat gactaggcat caggtacatg aaaaagaaga attcaaggct ttgaaaacgt
     5461 taagtatttt ctaccaagct gggacttcca aagctgggaa tcctattttt tattatgttg
     5521 cacggaggta agaaatacta tgttttgggt ctcttaacag aattttttaa attatagcaa
     5581 atatagagag atggcaacta gattagtggc atctgtatat ttattttaaa cactgctaat
     5641 aatctttgtc ttttttgtca ttttccttag gttcaaaact ggtcaaatca atggtgattt
     5701 gctgatatac catgtcttac tgactttaaa gccatattat gcaaagccat atgaaattgt
     5761 agtggacctt acccataccg ggcctagcaa tcgctttaaa acagactttc tctctaagtg
     5821 gtttgttgtt tttcctggct ttgcttacga caacgtctcc gcagtctata tctataactg
     5881 taactcctgg gtcagggagt acaccaagta tcatgagcgg ctgctgactg gcctcaaagg
     5941 tagcaaaagg cttgttttca tagactgtcc tgggaaactg gctgagcaca tagagcatga
     6001 acaacagaaa ctacctgctg ccaccttggc tttagaagag gacctgaagg tattccacaa
     6061 tgctctcaag ctagctcaca aagacaccaa agtttctatt aaagttggtt ctactgctgt
     6121 ccaagtaact tcagcagagc gaacaaaagt cctagggcaa tcagtctttc taaatgacat
     6181 ttattatgct tcggaaattg aagaaatctg cctagtagat gagaaccagt tcaccttaac
     6241 cattgcaaac cagggcacgc cgctcacctt catgcaccag gagtgtgaag ccattgtcca
     6301 gtctatcatt catatccgga cccgctggga actgtcacag cccgactcta tcccccaaca
     6361 caccaagatt cggccaaaag atgtccctgg gacactgctc aatatcgcat tacttaattt
     6421 aggcagttct gacccgagtt tacggtcagc tgcctataat cttctgtgtg ccttaacttg
     6481 tacctttaat ttaaaaatcg agggccagtt actagagaca tcaggtttat gtatccctgc
     6541 caacaacacc ctctttattg tctctattag taagacactg gcagccaatg agccacacct
     6601 cacgttagaa tttttggaag agtgtatttc tggatttagc aaatctagta ttgaattgaa
     6661 acacctttgt ttggaataca tgactccatg gctgtcaaat ctagttcgtt tttgcaagca
     6721 taatgatgat gccaaacgac aaagagttac tgctattctt gacaagctga taacaatgac
     6781 catcaatgaa aaacagatgt acccatctat tcaagcaaaa atatggggaa gccttgggca
     6841 gattacagat ctgcttgatg ttgtactaga cagtttcatc aaaaccagtg caacaggtgg
     6901 cttgggatca ataaaagctg aggtgatggc agatactgct gtagctttgg cttctggaaa
     6961 tgtgaaattg gtttcaagca aggttattgg aaggatgtgc aaaataattg acaagacatg
     7021 cttatctcca actcctactt tagaacaaca tcttatgtgg gatgatattg ctattttagc
     7081 acgctacatg ctgatgctgt ccttcaacaa ttcccttgat gtggcagctc atcttcccta
     7141 cctcttccac gttgttactt tcttagtagc cacaggtccg ctctccctta gagcttccac
     7201 acatggactg gtcattaata tcattcactc tctgtgtact tgttcacagc ttcattttag
     7261 tgaagagacc aagcaagttt tgagactcag tctgacagag ttctcattac ccaaatttta
     7321 cttgctgttt ggcattagca aagtcaagtc agctgctgtc attgccttcc gttccagtta
     7381 ccgggacagg tcattctctc ctggctccta tgagagagag acttttgctt tgacatcctt
     7441 ggaaacagtc acagaagctt tgttggagat catggaggca tgcatgagag atattccaac
     7501 gtgcaagtgg ctggaccagt ggacagaact agctcaaaga tttgcattcc aatataatcc
     7561 atccctgcaa ccaagagctc ttgttgtctt tgggtgtatt agcaaacgag tgtctcatgg
     7621 gcagataaag cagataatcc gtattcttag caaggcactt gagagttgct taaaaggacc
     7681 tgacacttac aacagtcaag ttctgataga agctacagta atagcactaa ccaaattaca
     7741 gccacttctt aataaggact cgcctctgca caaagccctc ttttgggtag ctgtggctgt
     7801 gctgcagctt gatgaggtca acttgtattc agcaggtacc gcacttcttg aacaaaacct
     7861 gcatacttta gatagtctcc gtatattcaa tgacaagagt ccagaggaag tatttatggc
     7921 aatccggaat cctctggagt ggcactgcaa gcaaatggat cattttgttg gactcaattt
     7981 caactctaac tttaactttg cattggttgg acacctttta aaagggtaca ggcatccttc
     8041 acctgctatt gttgcaagaa cagtcagaat tttacataca ctactaactc tggttaacaa
     8101 acacagaaat tgtgacaaat ttgaagtgaa tacacagagc gtggcctact tagcagcttt
     8161 acttacagtg tctgaagaag ttcgaagtcg ctgcagccta aaacatagaa agtcacttct
     8221 tcttactgat atttcaatgg aaaatgttcc tatggataca tatcccattc atcatggtga
     8281 cccttcctat aggacactaa aggagactca gccatggtcc tctcccaaag gttctgaagg
     8341 ataccttgca gccacctatc caactgtcgg ccagaccagt ccccgagcca ggaaatccat
     8401 gagcctggac atggggcaac cttctcaggc caacactaag aagttgcttg gaacaaggaa
     8461 aagttttgat cacttgatat cagacacaaa ggctcctaaa aggcaagaaa tggaatcagg
     8521 gatcacaaca ccccccaaaa tgaggagagt agcagaaact gattatgaaa tggaaactca
     8581 gaggatttcc tcatcacaac agcacccaca tttacgtaaa gtttcagtgt ctgaatcaaa
     8641 tgttctcttg gatgaagaag tacttactga tccgaagatc caggcgctgc ttcttactgt
     8701 tctagctaca ctggtaaaat ataccacaga tgagtttgat caacgaattc tttatgaata
     8761 cttagcagag gccagtgttg tgtttcccaa agtctttcct gttgtgcata atttgttgga
     8821 ctctaagatc aacaccctgt tatcattgtg ccaagatcca aatttgttaa atccaatcca
     8881 tggaattgtg cagagtgtgg tgtaccatga agaatcccca ccacaatacc aaacatctta
     8941 cctgcaaagt tttggtttta atggcttgtg gcggtttgca ggaccgtttt caaagcaaac
     9001 acaaattcca gactatgctg agcttattgt taagtttctt gatgccttga ttgacacgta
     9061 cctgcctgga attgatgaag aaaccagtga agaatccctc ctgactccca catctcctta
     9121 ccctcctgca ctgcagagcc agcttagtat cactgccaac cttaaccttt ctaattccat
     9181 gacctcactt gcaacttccc agcattcccc aggaatcgac aaggagaacg ttgaactctc
     9241 ccctaccact ggccactgta acagtggacg aactcgccac ggatccgcaa gccaagtgca
     9301 gaagcaaaga agcgctggca gtttcaaacg taatagcatt aagaagatcg tgtagcggcc
     9361 gcatcgtgac tgactgacga tctgcctcgc gcgtttcggt gatgacggtg aaaacctctg
     9421 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     9481 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc
     9541 acgtagcgat agcggagtgt ataattcttg aagacgaaag ggcctcgtga tacgcctatt
     9601 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     9661 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     9721 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     9781 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     9841 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     9901 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     9961 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
    10021 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
    10081 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
    10141 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
    10201 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
    10261 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc
    10321 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
    10381 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
    10441 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
    10501 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
    10561 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
    10621 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
    10681 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataa
//
