LOCUS       sequence-316755-        6283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP from the Gfa2 promoter. This plasmid is a new version 
            of Plasmid 22672 and includes a stop codon in the myrEGFP insert..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6283)
  AUTHORS   Nam HS, Benezra R
  TITLE     High levels of Id1 expression define B1 type adult neural stem 
            cells.
  JOURNAL   Cell Stem Cell. 2009 Nov 6. 5(5):515-26.
  PUBMED    19896442
REFERENCE   2  (bases 1 to 6283)
  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: "Cell Stem 
            Cell. 2009 Nov 6. 5(5):515-26."
COMMENT     Sequence Label: pGFAP::MyrGFP_v2
FEATURES             Location/Qualifiers
     source          1..6283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(3373..3375,3376..4089)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(3373..3375,3376..4086)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3415..3436)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3676..3695)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4023..4044
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4658..4675)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4658..4674)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4667..4689)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4883..4902)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5002..5024
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5062..5080)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5148..5252
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5253..5321,5322..6113)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga catgattacg aattcgagct cggtaccaga
     1021 tctgagctcc cacctccctc tctgtgctgg gactcacaga gggagacctc aggaggcagt
     1081 ctgtccatca catgtccaaa tgcagagcat accctgggct gggcgcagtg gcgcacaact
     1141 gtaattccag cactttggga ggctgatgtg gaaggatcac ttgagcccag aagttctaga
     1201 ccagcctggg caacatggca agaccctatc tctacaaaaa aagttaaaaa atcagccacg
     1261 tgtggtgaca cacacctgta gtcccagcta ttcaggaggc tgaggtgagg ggatcactta
     1321 aggctgggag gttgaggctg cagtgagtcg tggttgcgcc actgcactcc agcctgggca
     1381 acagtgagac cctgtctcaa aagacaaaaa aaaaaaaaaa aaaaaaaaga acatatcctg
     1441 gtgtggagta ggggacgctg ctctgacaga ggctcggggg cctgagctgg ctctgtgagc
     1501 tggggaggag gcagacagcc aggccttgtc tgcaagcaga cctggcagca ttgggctggc
     1561 cgccccccag ggcctcctct tcatgcccag tgaatgactc accttggcac agacacaatg
     1621 ttcggggtgg gcacagtgcc tgcttcccgc cgcaccccag cccccctcaa atgccttccg
     1681 agaagcccat tgagcagggg gcttgcattg caccccagcc tgacagcctg gcatcttggg
     1741 ataaaagcag cacagccccc taggggctgc ccttgctgtg tggcgccacc ggcggtggag
     1801 aacaaggctc tattcagcct gtgcccagga aaggggatca ggggatgccc aggcatggac
     1861 agtgggtggc agggggggag aggagggctg tctgcttccc agaagtccaa ggacacaaat
     1921 gggtgagggg actgggcagg gttctgaccc tgtgggacca gagtggaggg cgtagatgga
     1981 cctgaagtct ccagggacaa cagggcccag gtctcaggct cctagttggg cccagtggct
     2041 ccagcgtttc caaacccatc catccccaga ggttcttccc atctctccag gctgatgtgt
     2101 gggaactcga ggaaataaat ctccagtggg agacggaggg gtggccaggg aaacggggcg
     2161 ctgcaggaat aaagacgagc cagcacagcc agctcatgtg taacggcttt gtggagctgt
     2221 caaggcctgg tctctgggag agaggcacag ggaggccaga caaggaaggg gtgacctgga
     2281 gggacagatc caggggctaa agtcctgata aggcaagaga gtgccggccc cctcttgccc
     2341 tatcaggacc tccactgcca catagaggcc atgattgacc cttagacaaa gggctggtgt
     2401 ccaatcccag cccccagccc cagaactcca gggaatgaat gggcagagag caggaatgtg
     2461 ggacatctgt gttcaaggga aggactccag gagtctgctg ggaatgaggc ctagtaggaa
     2521 atgaggtggc ccttgagggt acagaacagg ttcattcttc gccaaattcc cagcaccttg
     2581 caggcactta cagctgagtg agataatgcc tgggttatga aatcaaaaag ttggaaagca
     2641 ggtcagaggt catctggtac agcccttcct tccctttttt tttttttttt ttgtgagaca
     2701 aggtctctct ctgttgccca ggctggagtg gcgcaaacac agctcactgc agcctcaacc
     2761 tactgggctc aagcaatcct ccagcctcag cctcccaaag tgctgggatt acaagcatga
     2821 gccaccccac tcagcccttt ccttcctttt taattgatgc ataataattg taagtattca
     2881 tcatggtcca accaaccctt tcttgaccca ccttcctaga gagagggtcc tcttgcttca
     2941 gcggtcaggg ccccagaccc atggtctggc tccaggtacc acctgcctca tgcaggagtt
     3001 ggcgtgccca ggaagctctg cctctgggca cagtgacctc agtggggtga ggggagctct
     3061 ccccatagct gggctgcggc ccaaccccac cccctcaggc tatgccaggg ggtgttgcca
     3121 ggggcacccg ggcatcgcca gtctagccca ctccttcata aagccctcgc atcccaggag
     3181 cgagcagagc cagagcaggt tggagaggag acgcatcacc tccgctgctc gcggggatcc
     3241 accatgggtt gctgtttctc caagaccgca gcgaagggag aagccgccgc ggagaggcct
     3301 ggggaggcgg ctgtggcctc gtcgccttcc aaagcgaacg gacaggagaa tggccacgtg
     3361 aaggtaggat ccgtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     3421 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     3481 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     3541 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     3601 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     3661 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     3721 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     3781 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     3841 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     3901 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     3961 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     4021 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     4081 tacaagtaag gatccgcaga tcccggccag ataccgatgc tgccgcagca aaagcaggag
     4141 cagatgccgc cgtcgcaggc gaagatgtcg cagacggagg aggcgatgct gccggcggag
     4201 gaggcgaagt aagtagaggg ctgggctggg ctgtgggggg tgtggggtgc gggactgggc
     4261 agtctgggag tccctctcac cacttttctt acctttctag gatgctgccg tcgccgccgc
     4321 tcatacacca taaggtgtaa aaaatactag atgcacagaa tagcaagtcc atcaaaactc
     4381 ctgcgtgaga attttaccag acttcaagag catctcgcca catcttgaaa aatgccaccg
     4441 tccgatgaaa aacaggagcc tgctaaggaa caatgccacc tgtcaataaa tgttgaaaac
     4501 tcatcccatt cctgcctctt ggtccttggg cttggggagg ggtgcgcgga tgtggttagg
     4561 gaacatgact ggtcaaatgg gaagggcttc aaaagaattc ccaatattga ctaccaagcc
     4621 acctgtacag atctgcctgc aggcatgcaa gcttggcact ggccgtcgtt ttacaacgtc
     4681 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     4741 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     4801 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     4861 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     4921 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     4981 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     5041 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     5101 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5161 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5221 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     5281 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5341 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5401 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5461 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5521 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5581 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5641 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5701 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5761 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     5821 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5881 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5941 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     6001 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     6061 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     6121 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6181 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6241 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttc
//
