LOCUS       sequence-229904-       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, KDR)
FEATURES             Location/Qualifiers
     source          1..10725
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     449..468
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(9296..9318)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9463..9481)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9549..9653
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9654..9722,9723..10514)
                     /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(9872..9891)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(10685..10725,1..548)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
       61 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
      121 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
      181 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
      241 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
      301 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
      361 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
      421 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
      481 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
      541 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
      601 ctcacatgtc gtacgacgcg tcccgggacc ggtggcatgg ccgcgcacag gccggtggaa
      661 tgggtccagg ccgtggtcag ccgcttcgac gagcagcttc caataaaaac aggacagcag
      721 aacacacata ccaaagtcag tactgagcac aacaaggaat gtctaatcaa tatttccaaa
      781 tacaagtttt ctttggttat aagcggcctc actactattt taaagaatgt taacaatatg
      841 agaatatttg gagaagctgc tgaaaaaaat ttatatctct ctcagttgat tatattggat
      901 acactggaaa aatgtcttgc tgggcaacca aaggacacaa tgagattaga tgaaacgatg
      961 ctggtcaaac agttgctgcc agaaatctgc cattttcttc acacctgtcg tgaaggaaac
     1021 cagcatgcag ctgaacttcg gaattctgcc tctggggttt tattttctct cagctgcaac
     1081 aacttcaatg cagtctttag tcgcatttct accaggttac aggaattaac tgtttgttca
     1141 gaagacaatg ttgatgttca tgatatagaa ttgttacagt atatcaatgt ggattgtgca
     1201 aaattaaaac gactcctgaa ggaaacagca tttaaattta aagccctaaa gaaggttgcg
     1261 cagttagcag ttataaatag cctggaaaag gcattttgga actgggtaga aaattatcca
     1321 gatgaattta caaaactata ccagatccca cagactgata tggctgaatg tgcagaaaag
     1381 ctatttgact tggtggatgg ttttgctgaa agcaccaaac gtaaagcagc agtttggcca
     1441 ctacaaatca ttctccttat cttgtgtcca gaaataatcc aggatatatc caaagacgtg
     1501 gttgatgaaa acaacatgaa taagaagtta tttctggaca gtctacgaaa agctcttgct
     1561 ggccatggag gaagtaggca gctgacagaa agtgctgcaa ttgcctgtgt caaactgtgt
     1621 aaagcaagta cttacatcaa ttgggaagat aactctgtca ttttcctact tgttcagtcc
     1681 atggtggttg atcttaagaa cctgcttttt aatccaagta agccattctc aagaggcagt
     1741 cagcctgcag atgtggatct aatgattgac tgccttgttt cttgctttcg tataagccct
     1801 cacaacaacc aacactttaa gatctgcctg gctcagaatt caccttctac atttcactat
     1861 gtgctggtaa attcactcca tcgaatcatc accaattccg cattggattg gtggcctaag
     1921 attgatgctg tgtattgtca ctcggttgaa cttcgaaata tgtttggtga aacacttcat
     1981 aaagcagtgc aaggttgtgg agcacaccca gcaatacgaa tggcaccgag tcttacattt
     2041 aaagaaaaag taacaagcct taaatttaaa gaaaaaccta cagacctgga gacaagaagc
     2101 tataagtatc ttctcttgtc catggtgaaa ctaattcatg cagatccaaa gctcttgctt
     2161 tgtaatccaa gaaaacaggg gcccgaaacc caaggcagta cagcagaatt aattacaggg
     2221 ctcgtccaac tggtccctca gtcacacatg ccagagattg ctcaggaagc aatggaggct
     2281 ctgctggttc ttcatcagtt agatagcatt gatttgtgga atcctgatgc tcctgtagaa
     2341 acattttggg agattagctc acaaatgctt ttttacatct gcaagaaatt aactagtcat
     2401 caaatgctta gtagcacaga aattctcaag tggttgcggg aaatattgat ctgcaggaat
     2461 aaatttcttc ttaaaaataa gcaggcagat agaagttcct gtcactttct ccttttttac
     2521 ggggtaggat gtgatattcc ttctagtgga aataccagtc aaatgtccat ggatcatgaa
     2581 gaattactac gtactcctgg agcctctctc cggaagggaa aagggaactc ctctatggat
     2641 agtgcagcag gatgcagcgg aaccccccca atttgccgac aagcccagac caaactagaa
     2701 gtggccctgt acatgtttct gtggaaccct gacactgaag ctgttctggt tgccatgtcc
     2761 tgtttccgcc acctctgtga ggaagcagat atccggtgtg gggtggatga agtgtcagtg
     2821 cataacctct tgcccaacta taacacattc atggagtttg cctctgtcag caatatgatg
     2881 tcaacaggaa gagcagcact tcagaaaaga gtgatggcac tgctgaggcg cattgagcat
     2941 cccactgcag gaaacactga ggcttgggaa gatacacatg caaaatggga acaagcaaca
     3001 aagctaatcc ttaactatcc aaaagccaaa atggaagatg gccaggctgc tgaaagcctt
     3061 cacaagacca ttgtcaagag gcgaatgtcc catgtgagtg gaggaggatc catagatttg
     3121 tctgacacag actccctaca ggaatggatc aacatgactg gcttcctttg tgcccttggg
     3181 ggagtgtgcc tccagcagag aagcaattct ggcctggcaa cctatagccc acccatgggt
     3241 ccagtcagtg aacgtaaggg ttctatgatt tcagtgatgt cttcagaggg aaacgcagat
     3301 acacctgtca gcaaatttat ggatcggctg ttgtccttaa tggtgtgtaa ccatgagaaa
     3361 gtgggacttc aaatacggac caatgttaag gatctggtgg gtctagaatt gagtcctgct
     3421 ctgtatccaa tgctatttaa caaattgaag aataccatca gcaagttttt tgactcccaa
     3481 ggacaggttt tattgactga taccaatact caatttgtag aacaaaccat agctataatg
     3541 aagaacttgc tagataatca tactgaaggc agctctgaac atctagggca agctagcatt
     3601 gaaacaatga tgttaaatct ggtcaggtat gttcgtgtgc ttgggaatat ggtccatgca
     3661 attcaaataa aaacgaaact gtgtcaatta gttgaagtaa tgatggcaag gagagatgac
     3721 ctctcatttt gccaagagat gaaatttagg aataagatgg tagaatacct gacagactgg
     3781 gttatgggaa catcaaacca agcagcagat gatgatgtaa aatgtcttac aagagatttg
     3841 gaccaggcaa gcatggaagc agtagtttca cttctagctg gtctccctct gcagcctgaa
     3901 gaaggagatg gtgtggaatt gatggaagcc aaatcacagt tatttcttaa atacttcaca
     3961 ttatttatga accttttgaa tgactgcagt gaagttgaag atgaaagtgc gcaaacaggt
     4021 ggcaggaaac gtggcatgtc tcggaggctg gcatcactga ggcactgtac ggtccttgca
     4081 atgtcaaact tactcaatgc caacgtagac agtggtctca tgcactccat aggcttaggt
     4141 taccacaagg atctccagac aagagctaca tttatggaag ttctgacaaa aatccttcaa
     4201 caaggcacag aatttgacac acttgcagaa acagtattgg ctgatcggtt tgagagattg
     4261 gtggaactgg tcacaatgat gggtgatcaa ggagaactcc ctatagcgat ggctctggcc
     4321 aatgtggttc cttgttctca gtgggatgaa ctagctcgag ttctggttac tctgtttgat
     4381 tctcggcatt tactctacca actgctctgg aacatgtttt ctaaagaagt agaattggca
     4441 gactccatgc agactctctt ccgaggcaac agcttggcca gtaaaataat gacattctgt
     4501 ttcaaggtat atggtgctac ctatctacaa aaactcctgg atcctttatt acgaattgtg
     4561 atcacatcct ctgattggca acatgttagc tttgaagtgg atcctaccag gttagaacca
     4621 tcagagagcc ttgaggaaaa ccagcggaac ctccttcaga tgactgaaaa gttcttccat
     4681 gccatcatca gttcctcctc agaattcccc cctcaacttc gaagtgtgtg ccactgttta
     4741 taccaggtgg ttagccagcg tttccctcag aacagcatcg gtgcagtagg aagtgccatg
     4801 ttcctcagat ttatcaatcc tgccattgtc tcaccgtatg aagcagggat tttagataaa
     4861 aagccaccac ctagaatcga aaggggcttg aagttaatgt caaagatact tcagagtatt
     4921 gccaatcatg ttctcttcac aaaagaagaa catatgcggc ctttcaatga ttttgtgaaa
     4981 agcaactttg atgcagcacg caggtttttc cttgatatag catctgattg tcctacaagt
     5041 gatgcagtaa atcatagtct ttccttcata agtgacggca atgtgcttgc tttacatcgt
     5101 ctactctgga acaatcagga gaaaattggg cagtatcttt ccagcaacag ggatcataaa
     5161 gctgttggaa gacgaccttt tgataagatg gcaacacttc ttgcatacct gggtcctcca
     5221 gagcacaaac ctgtggcaga tacacactgg tccagcctta accttaccag ttcaaagttt
     5281 gaggaattta tgactaggca tcaggtacat gaaaaagaag aattcaaggc tttgaaaacg
     5341 ttaagtattt tctaccaagc tgggacttcc aaagctggga atcctatttt ttattatgtt
     5401 gcacggaggt aagaaatact atgttttggg tctcttaaca gaatttttta aattatagca
     5461 aatatagaga gatggcaact agattagtgg catctgtata tttattttaa acactgctaa
     5521 taatctttgt cttttttgtc attttcctta ggttcaaaac tggtcaaatc aatggtgatt
     5581 tgctgatata ccatgtctta ctgactttaa agccatatta tgcaaagcca tatgaaattg
     5641 tagtggacct tacccatacc gggcctagca atcgctttaa aacagacttt ctctctaagt
     5701 ggtttgttgt ttttcctggc tttgcttacg acaacgtctc cgcagtctat atctataact
     5761 gtaactcctg ggtcagggag tacaccaagt atcatgagcg gctgctgact ggcctcaaag
     5821 gtagcaaaag gcttgttttc atagactgtc ctgggaaact ggctgagcac atagagcatg
     5881 aacaacagaa actacctgct gccaccttgg ctttagaaga ggacctgaag gtattccaca
     5941 atgctctcaa gctagctcac aaagacacca aagtttctat taaagttggt tctactgctg
     6001 tccaagtaac ttcagcagag cgaacaaaag tcctagggca atcagtcttt ctaaatgaca
     6061 tttattatgc ttcggaaatt gaagaaatct gcctagtaga tgagaaccag ttcaccttaa
     6121 ccattgcaaa ccagggcacg ccgctcacct tcatgcacca ggagtgtgaa gccattgtcc
     6181 agtctatcat tcatatccgg acccgctggg aactgtcaca gcccgactct atcccccaac
     6241 acaccaagat tcggccaaaa gatgtccctg ggacactgct caatatcgca ttacttaatt
     6301 taggcagttc tgacccgagt ttacggtcag ctgcctataa tcttctgtgt gccttaactt
     6361 gtacctttaa tttaaaaatc gagggccagt tactagagac atcaggttta tgtatccctg
     6421 ccaacaacac cctctttatt gtctctatta gtaagacact ggcagccaat gagccacacc
     6481 tcacgttaga atttttggaa gagtgtattt ctggatttag caaatctagt attgaattga
     6541 aacacctttg tttggaatac atgactccat ggctgtcaaa tctagttcgt ttttgcaagc
     6601 ataatgatga tgccaaacga caaagagtta ctgctattct tgacaagctg ataacaatga
     6661 ccatcaatga aaaacagatg tacccatcta ttcaagcaaa aatatgggga agccttgggc
     6721 agattacaga tctgcttgat gttgtactag acagtttcat caaaaccagt gcaacaggtg
     6781 gcttgggatc aataaaagct gaggtgatgg cagatactgc tgtagctttg gcttctggaa
     6841 atgtgaaatt ggtttcaagc aaggttattg gaaggatgtg caaaataatt gacaagacat
     6901 gcttatctcc aactcctact ttagaacaac atcttatgtg ggatgatatt gctattttag
     6961 cacgctacat gctgatgctg tccttcaaca attcccttga tgtggcagct catcttccct
     7021 acctcttcca cgttgttact ttcttagtag ccacaggtcc gctctccctt agagcttcca
     7081 cacatggact ggtcattaat atcattcact ctctgtgtac ttgttcacag cttcatttta
     7141 gtgaagagac caagcaagtt ttgagactca gtctgacaga gttctcatta cccaaatttt
     7201 acttgctgtt tggcattagc aaagtcaagt cagctgctgt cattgccttc cgttccagtt
     7261 accgggacag gtcattctct cctggctcct atgagagaga gacttttgct ttgacatcct
     7321 tggaaacagt cacagaagct ttgttggaga tcatggaggc atgcatgaga gatattccaa
     7381 cgtgcaagtg gctggaccag tggacagaac tagctcaaag atttgcattc caatataatc
     7441 catccctgca accaagagct cttgttgtct ttgggtgtat tagcaaacga gtgtctcatg
     7501 ggcagataaa gcagataatc cgtattctta gcaaggcact tgagagttgc ttaaaaggac
     7561 ctgacactta caacagtcaa gttctgatag aagctacagt aatagcacta accaaattac
     7621 agccacttct taataaggac tcgcctctgc acaaagccct cttttgggta gctgtggctg
     7681 tgctgcagct tgatgaggtc aacttgtatt cagcaggtac cgcacttctt gaacaaaacc
     7741 tgcatacttt agatagtctc cgtatattca atgacaagag tccagaggaa gtatttatgg
     7801 caatccggaa tcctctggag tggcactgca agcaaatgga tcattttgtt ggactcaatt
     7861 tcaactctaa ctttaacttt gcattggttg gacacctttt aaaagggtac aggcatcctt
     7921 cacctgctat tgttgcaaga acagtcagaa ttttacatac actactaact ctggttaaca
     7981 aacacagaaa ttgtgacaaa tttgaagtga atacacagag cgtggcctac ttagcagctt
     8041 tacttacagt gtctgaagaa gttcgaagtc gctgcagcct aaaacataga aagtcacttc
     8101 ttcttactga tatttcaatg gaaaatgttc ctatggatac atatcccatt catcatggtg
     8161 acccttccta taggacacta aaggagactc agccatggtc ctctcccaaa ggttctgaag
     8221 gataccttgc agccacctat ccaactgtcg gccagaccag tccccgagcc aggaaatcca
     8281 tgagcctgga catggggcaa ccttctcagg ccaacactaa gaagttgctt ggaacaagga
     8341 aaagttttga tcacttgata tcagacacaa aggctcctaa aaggcaagaa atggaatcag
     8401 ggatcacaac accccccaaa atgaggagag tagcagaaac tgattatgaa atggaaactc
     8461 agaggatttc ctcatcacaa cagcacccac atttacgtaa agtttcagtg tctgaatcaa
     8521 atgttctctt ggatgaagaa gtacttactg atccgaagat ccaggcgctg cttcttactg
     8581 ttctagctac actggtaaaa tataccacag atgagtttga tcaacgaatt ctttatgaat
     8641 acttagcaga ggccagtgtt gtgtttccca aagtctttcc tgttgtgcat aatttgttgg
     8701 actctaagat caacaccctg ttatcattgt gccaagatcc aaatttgtta aatccaatcc
     8761 atggaattgt gcagagtgtg gtgtaccatg aagaatcccc accacaatac caaacatctt
     8821 acctgcaaag ttttggtttt aatggcttgt ggcggtttgc aggaccgttt tcaaagcaaa
     8881 cacaaattcc agactatgct gagcttattg ttaagtttct tgatgccttg attgacacgt
     8941 acctgcctgg aattgatgaa gaaaccagtg aagaatccct cctgactccc acatctcctt
     9001 accctcctgc actgcagagc cagcttagta tcactgccaa ccttaacctt tctaattcca
     9061 tgacctcact tgcaacttcc cagcattccc caggaatcga caaggagaac gttgaactct
     9121 cccctaccac tggccactgt aacagtggac gaactcgcca cggatccgca agccaagtgc
     9181 agaagcaaag aagcgcagga tcatttaaga gaaatagcat taagaagatc gtgtagcggc
     9241 cgcatcgtga ctgactgacg atctgcctcg cgcgtttcgg tgatgacggt gaaaacctct
     9301 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     9361 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt
     9421 cacgtagcga tagcggagtg tataattctt gaagacgaaa gggcctcgtg atacgcctat
     9481 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     9541 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     9601 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     9661 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     9721 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     9781 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     9841 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
     9901 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     9961 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
    10021 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
    10081 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
    10141 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag
    10201 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
    10261 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
    10321 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
    10381 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
    10441 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
    10501 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
    10561 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
    10621 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
    10681 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaa
//
