LOCUS       sequence-231644-       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 R1276P (+minintr, KDR)
FEATURES             Location/Qualifiers
     source          1..10725
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      668..1256
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1157..1176
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(10004..10026)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(10171..10189)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        10257..10361
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10362..10430,10431..10725,1..497)
                     /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(10580..10599)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
       61 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
      121 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
      181 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
      241 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
      301 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
      361 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
      421 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
      481 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
      541 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
      601 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
      661 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
      721 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
      781 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
      841 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
      901 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
      961 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     1021 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     1081 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     1141 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     1201 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     1261 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtcgt
     1321 acgacgcgtc ccgggaccgg tggcatggcc gcgcacaggc cggtggaatg ggtccaggcc
     1381 gtggtcagcc gcttcgacga gcagcttcca ataaaaacag gacagcagaa cacacatacc
     1441 aaagtcagta ctgagcacaa caaggaatgt ctaatcaata tttccaaata caagttttct
     1501 ttggttataa gcggcctcac tactatttta aagaatgtta acaatatgag aatatttgga
     1561 gaagctgctg aaaaaaattt atatctctct cagttgatta tattggatac actggaaaaa
     1621 tgtcttgctg ggcaaccaaa ggacacaatg agattagatg aaacgatgct ggtcaaacag
     1681 ttgctgccag aaatctgcca ttttcttcac acctgtcgtg aaggaaacca gcatgcagct
     1741 gaacttcgga attctgcctc tggggtttta ttttctctca gctgcaacaa cttcaatgca
     1801 gtctttagtc gcatttctac caggttacag gaattaactg tttgttcaga agacaatgtt
     1861 gatgttcatg atatagaatt gttacagtat atcaatgtgg attgtgcaaa attaaaacga
     1921 ctcctgaagg aaacagcatt taaatttaaa gccctaaaga aggttgcgca gttagcagtt
     1981 ataaatagcc tggaaaaggc attttggaac tgggtagaaa attatccaga tgaatttaca
     2041 aaactatacc agatcccaca gactgatatg gctgaatgtg cagaaaagct atttgacttg
     2101 gtggatggtt ttgctgaaag caccaaacgt aaagcagcag tttggccact acaaatcatt
     2161 ctccttatct tgtgtccaga aataatccag gatatatcca aagacgtggt tgatgaaaac
     2221 aacatgaata agaagttatt tctggacagt ctacgaaaag ctcttgctgg ccatggagga
     2281 agtaggcagc tgacagaaag tgctgcaatt gcctgtgtca aactgtgtaa agcaagtact
     2341 tacatcaatt gggaagataa ctctgtcatt ttcctacttg ttcagtccat ggtggttgat
     2401 cttaagaacc tgctttttaa tccaagtaag ccattctcaa gaggcagtca gcctgcagat
     2461 gtggatctaa tgattgactg ccttgtttct tgctttcgta taagccctca caacaaccaa
     2521 cactttaaga tctgcctggc tcagaattca ccttctacat ttcactatgt gctggtaaat
     2581 tcactccatc gaatcatcac caattccgca ttggattggt ggcctaagat tgatgctgtg
     2641 tattgtcact cggttgaact tcgaaatatg tttggtgaaa cacttcataa agcagtgcaa
     2701 ggttgtggag cacacccagc aatacgaatg gcaccgagtc ttacatttaa agaaaaagta
     2761 acaagcctta aatttaaaga aaaacctaca gacctggaga caagaagcta taagtatctt
     2821 ctcttgtcca tggtgaaact aattcatgca gatccaaagc tcttgctttg taatccaaga
     2881 aaacaggggc ccgaaaccca aggcagtaca gcagaattaa ttacagggct cgtccaactg
     2941 gtccctcagt cacacatgcc agagattgct caggaagcaa tggaggctct gctggttctt
     3001 catcagttag atagcattga tttgtggaat cctgatgctc ctgtagaaac attttgggag
     3061 attagctcac aaatgctttt ttacatctgc aagaaattaa ctagtcatca aatgcttagt
     3121 agcacagaaa ttctcaagtg gttgcgggaa atattgatct gcaggaataa atttcttctt
     3181 aaaaataagc aggcagatag aagttcctgt cactttctcc ttttttacgg ggtaggatgt
     3241 gatattcctt ctagtggaaa taccagtcaa atgtccatgg atcatgaaga attactacgt
     3301 actcctggag cctctctccg gaagggaaaa gggaactcct ctatggatag tgcagcagga
     3361 tgcagcggaa cccccccaat ttgccgacaa gcccagacca aactagaagt ggccctgtac
     3421 atgtttctgt ggaaccctga cactgaagct gttctggttg ccatgtcctg tttccgccac
     3481 ctctgtgagg aagcagatat ccggtgtggg gtggatgaag tgtcagtgca taacctcttg
     3541 cccaactata acacattcat ggagtttgcc tctgtcagca atatgatgtc aacaggaaga
     3601 gcagcacttc agaaaagagt gatggcactg ctgaggcgca ttgagcatcc cactgcagga
     3661 aacactgagg cttgggaaga tacacatgca aaatgggaac aagcaacaaa gctaatcctt
     3721 aactatccaa aagccaaaat ggaagatggc caggctgctg aaagccttca caagaccatt
     3781 gtcaagaggc gaatgtccca tgtgagtgga ggaggatcca tagatttgtc tgacacagac
     3841 tccctacagg aatggatcaa catgactggc ttcctttgtg cccttggggg agtgtgcctc
     3901 cagcagagaa gcaattctgg cctggcaacc tatagcccac ccatgggtcc agtcagtgaa
     3961 cgtaagggtt ctatgatttc agtgatgtct tcagagggaa acgcagatac acctgtcagc
     4021 aaatttatgg atcggctgtt gtccttaatg gtgtgtaacc atgagaaagt gggacttcaa
     4081 atacggacca atgttaagga tctggtgggt ctagaattga gtcctgctct gtatccaatg
     4141 ctatttaaca aattgaagaa taccatcagc aagttttttg actcccaagg acaggtttta
     4201 ttgactgata ccaatactca atttgtagaa caaaccatag ctataatgaa gaacttgcta
     4261 gataatcata ctgaaggcag ctctgaacat ctagggcaag ctagcattga aacaatgatg
     4321 ttaaatctgg tcaggtatgt tcgtgtgctt gggaatatgg tccatgcaat tcaaataaaa
     4381 acgaaactgt gtcaattagt tgaagtaatg atggcaagga gagatgacct ctcattttgc
     4441 caagagatga aatttaggaa taagatggta gaatacctga cagactgggt tatgggaaca
     4501 tcaaaccaag cagcagatga tgatgtaaaa tgtcttacaa gagatttgga ccaggcaagc
     4561 atggaagcag tagtttcact tctagctggt ctccctctgc agcctgaaga aggagatggt
     4621 gtggaattga tggaagccaa atcacagtta tttcttaaat acttcacatt atttatgaac
     4681 cttttgaatg actgcagtga agttgaagat gaaagtgcgc aaacaggtgg caggaaacgt
     4741 ggcatgtctc ggaggctggc atcactgagg cactgtacgg tccttgcaat gtcaaactta
     4801 ctcaatgcca acgtagacag tggtctcatg cactccatag gcttaggtta ccacaaggat
     4861 ctccagacaa gagctacatt tatggaagtt ctgacaaaaa tccttcaaca aggcacagaa
     4921 tttgacacac ttgcagaaac agtattggct gatcggtttg agagattggt ggaactggtc
     4981 acaatgatgg gtgatcaagg agaactccct atagcgatgg ctctggccaa tgtggttcct
     5041 tgttctcagt gggatgaact agctcgagtt ctggttactc tgtttgattc tcggcattta
     5101 ctctaccaac tgctctggaa catgttttct aaagaagtag aattggcaga ctccatgcag
     5161 actctcttcc caggcaacag cttggccagt aaaataatga cattctgttt caaggtatat
     5221 ggtgctacct atctacaaaa actcctggat cctttattac gaattgtgat cacatcctct
     5281 gattggcaac atgttagctt tgaagtggat cctaccaggt tagaaccatc agagagcctt
     5341 gaggaaaacc agcggaacct ccttcagatg actgaaaagt tcttccatgc catcatcagt
     5401 tcctcctcag aattcccccc tcaacttcga agtgtgtgcc actgtttata ccaggtggtt
     5461 agccagcgtt tccctcagaa cagcatcggt gcagtaggaa gtgccatgtt cctcagattt
     5521 atcaatcctg ccattgtctc accgtatgaa gcagggattt tagataaaaa gccaccacct
     5581 agaatcgaaa ggggcttgaa gttaatgtca aagatacttc agagtattgc caatcatgtt
     5641 ctcttcacaa aagaagaaca tatgcggcct ttcaatgatt ttgtgaaaag caactttgat
     5701 gcagcacgca ggtttttcct tgatatagca tctgattgtc ctacaagtga tgcagtaaat
     5761 catagtcttt ccttcataag tgacggcaat gtgcttgctt tacatcgtct actctggaac
     5821 aatcaggaga aaattgggca gtatctttcc agcaacaggg atcataaagc tgttggaaga
     5881 cgaccttttg ataagatggc aacacttctt gcatacctgg gtcctccaga gcacaaacct
     5941 gtggcagata cacactggtc cagccttaac cttaccagtt caaagtttga ggaatttatg
     6001 actaggcatc aggtacatga aaaagaagaa ttcaaggctt tgaaaacgtt aagtattttc
     6061 taccaagctg ggacttccaa agctgggaat cctatttttt attatgttgc acggaggtaa
     6121 gaaatactat gttttgggtc tcttaacaga attttttaaa ttatagcaaa tatagagaga
     6181 tggcaactag attagtggca tctgtatatt tattttaaac actgctaata atctttgtct
     6241 tttttgtcat tttccttagg ttcaaaactg gtcaaatcaa tggtgatttg ctgatatacc
     6301 atgtcttact gactttaaag ccatattatg caaagccata tgaaattgta gtggacctta
     6361 cccataccgg gcctagcaat cgctttaaaa cagactttct ctctaagtgg tttgttgttt
     6421 ttcctggctt tgcttacgac aacgtctccg cagtctatat ctataactgt aactcctggg
     6481 tcagggagta caccaagtat catgagcggc tgctgactgg cctcaaaggt agcaaaaggc
     6541 ttgttttcat agactgtcct gggaaactgg ctgagcacat agagcatgaa caacagaaac
     6601 tacctgctgc caccttggct ttagaagagg acctgaaggt attccacaat gctctcaagc
     6661 tagctcacaa agacaccaaa gtttctatta aagttggttc tactgctgtc caagtaactt
     6721 cagcagagcg aacaaaagtc ctagggcaat cagtctttct aaatgacatt tattatgctt
     6781 cggaaattga agaaatctgc ctagtagatg agaaccagtt caccttaacc attgcaaacc
     6841 agggcacgcc gctcaccttc atgcaccagg agtgtgaagc cattgtccag tctatcattc
     6901 atatccggac ccgctgggaa ctgtcacagc ccgactctat cccccaacac accaagattc
     6961 ggccaaaaga tgtccctggg acactgctca atatcgcatt acttaattta ggcagttctg
     7021 acccgagttt acggtcagct gcctataatc ttctgtgtgc cttaacttgt acctttaatt
     7081 taaaaatcga gggccagtta ctagagacat caggtttatg tatccctgcc aacaacaccc
     7141 tctttattgt ctctattagt aagacactgg cagccaatga gccacacctc acgttagaat
     7201 ttttggaaga gtgtatttct ggatttagca aatctagtat tgaattgaaa cacctttgtt
     7261 tggaatacat gactccatgg ctgtcaaatc tagttcgttt ttgcaagcat aatgatgatg
     7321 ccaaacgaca aagagttact gctattcttg acaagctgat aacaatgacc atcaatgaaa
     7381 aacagatgta cccatctatt caagcaaaaa tatggggaag ccttgggcag attacagatc
     7441 tgcttgatgt tgtactagac agtttcatca aaaccagtgc aacaggtggc ttgggatcaa
     7501 taaaagctga ggtgatggca gatactgctg tagctttggc ttctggaaat gtgaaattgg
     7561 tttcaagcaa ggttattgga aggatgtgca aaataattga caagacatgc ttatctccaa
     7621 ctcctacttt agaacaacat cttatgtggg atgatattgc tattttagca cgctacatgc
     7681 tgatgctgtc cttcaacaat tcccttgatg tggcagctca tcttccctac ctcttccacg
     7741 ttgttacttt cttagtagcc acaggtccgc tctcccttag agcttccaca catggactgg
     7801 tcattaatat cattcactct ctgtgtactt gttcacagct tcattttagt gaagagacca
     7861 agcaagtttt gagactcagt ctgacagagt tctcattacc caaattttac ttgctgtttg
     7921 gcattagcaa agtcaagtca gctgctgtca ttgccttccg ttccagttac cgggacaggt
     7981 cattctctcc tggctcctat gagagagaga cttttgcttt gacatccttg gaaacagtca
     8041 cagaagcttt gttggagatc atggaggcat gcatgagaga tattccaacg tgcaagtggc
     8101 tggaccagtg gacagaacta gctcaaagat ttgcattcca atataatcca tccctgcaac
     8161 caagagctct tgttgtcttt gggtgtatta gcaaacgagt gtctcatggg cagataaagc
     8221 agataatccg tattcttagc aaggcacttg agagttgctt aaaaggacct gacacttaca
     8281 acagtcaagt tctgatagaa gctacagtaa tagcactaac caaattacag ccacttctta
     8341 ataaggactc gcctctgcac aaagccctct tttgggtagc tgtggctgtg ctgcagcttg
     8401 atgaggtcaa cttgtattca gcaggtaccg cacttcttga acaaaacctg catactttag
     8461 atagtctccg tatattcaat gacaagagtc cagaggaagt atttatggca atccggaatc
     8521 ctctggagtg gcactgcaag caaatggatc attttgttgg actcaatttc aactctaact
     8581 ttaactttgc attggttgga caccttttaa aagggtacag gcatccttca cctgctattg
     8641 ttgcaagaac agtcagaatt ttacatacac tactaactct ggttaacaaa cacagaaatt
     8701 gtgacaaatt tgaagtgaat acacagagcg tggcctactt agcagcttta cttacagtgt
     8761 ctgaagaagt tcgaagtcgc tgcagcctaa aacatagaaa gtcacttctt cttactgata
     8821 tttcaatgga aaatgttcct atggatacat atcccattca tcatggtgac ccttcctata
     8881 ggacactaaa ggagactcag ccatggtcct ctcccaaagg ttctgaagga taccttgcag
     8941 ccacctatcc aactgtcggc cagaccagtc cccgagccag gaaatccatg agcctggaca
     9001 tggggcaacc ttctcaggcc aacactaaga agttgcttgg aacaaggaaa agttttgatc
     9061 acttgatatc agacacaaag gctcctaaaa ggcaagaaat ggaatcaggg atcacaacac
     9121 cccccaaaat gaggagagta gcagaaactg attatgaaat ggaaactcag aggatttcct
     9181 catcacaaca gcacccacat ttacgtaaag tttcagtgtc tgaatcaaat gttctcttgg
     9241 atgaagaagt acttactgat ccgaagatcc aggcgctgct tcttactgtt ctagctacac
     9301 tggtaaaata taccacagat gagtttgatc aacgaattct ttatgaatac ttagcagagg
     9361 ccagtgttgt gtttcccaaa gtctttcctg ttgtgcataa tttgttggac tctaagatca
     9421 acaccctgtt atcattgtgc caagatccaa atttgttaaa tccaatccat ggaattgtgc
     9481 agagtgtggt gtaccatgaa gaatccccac cacaatacca aacatcttac ctgcaaagtt
     9541 ttggttttaa tggcttgtgg cggtttgcag gaccgttttc aaagcaaaca caaattccag
     9601 actatgctga gcttattgtt aagtttcttg atgccttgat tgacacgtac ctgcctggaa
     9661 ttgatgaaga aaccagtgaa gaatccctcc tgactcccac atctccttac cctcctgcac
     9721 tgcagagcca gcttagtatc actgccaacc ttaacctttc taattccatg acctcacttg
     9781 caacttccca gcattcccca ggaatcgaca aggagaacgt tgaactctcc cctaccactg
     9841 gccactgtaa cagtggacga actcgccacg gatccgcaag ccaagtgcag aagcaaagaa
     9901 gcgcaggatc atttaagaga aatagcatta agaagatcgt gtagcggccg catcgtgact
     9961 gactgacgat ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
    10021 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
    10081 gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata
    10141 gcggagtgta taattcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta
    10201 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
    10261 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
    10321 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
    10381 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
    10441 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
    10501 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
    10561 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
    10621 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
    10681 cagaaaagca tcttacggat ggcatgacag taagagaatt atgca
//
