LOCUS       pCAG-sfGFP                6569 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  sfGFP without iCreERT2 fusion.
ACCESSION   .
VERSION     .
KEYWORDS    pCAG-sfGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6569)
  AUTHORS   Nam HS, Capecchi MR
  TITLE     Lrig1 expression prospectively identifies stem cells in the 
            ventricular-subventricular zone that are neurogenic throughout adult
            life.
  JOURNAL   Neural Dev. 2020 Mar 17;15(1):3. doi: 10.1186/s13064-020-00139-5.
  PUBMED    32183906
REFERENCE   2  (bases 1 to 6569)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            //gt12365.cn
FEATURES             Location/Qualifiers
     source          1..6569
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             129..989
                     /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"
     primer_bind     complement(347..366)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1649..1668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1902..1919
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2072..2102
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /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     2115..2137
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2134..2150
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        2406..2785
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2787..3066
                     /label=chicken beta-actin promoter
     intron          3067..4083
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     4007..4029
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     4091..4110
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             4152..4865
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(4182..4210)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4805..4825
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4926..4946)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5344..5355)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5500..5517)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5506..5730
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5927..5946)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5928..5946)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5953..5970)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5953..5969)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5962..5984)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(6111..6566)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6248..6269)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6460..6479
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
       61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
      841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
      901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
      961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     1921 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     1981 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     2041 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     2101 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     2161 ccaagcgcgc aattaagttt aaacaagggc aggatgtgtc aaactgccaa tagagaacta
     2221 cttactcttc aggctgaagc tgatggaaca ggtaacaaag gcaaacacta atcatgatca
     2281 gcaagatgaa gcagaaaggg aacaagggga tattaaatgt gtatagacac gctagagaga
     2341 tggctcagca gttaagagaa ctagctggtc tttcagaggt cctgagatca ataccggtca
     2401 tggtcgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     2461 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     2521 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     2581 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     2641 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     2701 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     2761 cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc
     2821 catctccccc ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc
     2881 agcgatgggg gcgggggggg gggggggggg cgcgcgccag gcggggcggg gcggggcgag
     2941 gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga
     3001 aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg
     3061 cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc
     3121 cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct
     3181 tctcctccgg gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg
     3241 cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggggt
     3301 gcgtgcgtgt gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt
     3361 gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg
     3421 gccgggggcg gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg
     3481 ggtgtgtgcg tgggggggtg agcagggggt gtgggcgcgt cggtcgggct gcaacccccc
     3541 ctgcaccccc ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtacg
     3601 gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc
     3661 ggggcggggc cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg cccccggagc
     3721 gccggcggct gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag
     3781 agggcgcagg gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc
     3841 cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc
     3901 ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc
     3961 tgtccgcggg gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg
     4021 cgtgtgaccg gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta
     4081 cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattgcta
     4141 gcgccgccac catgagcaaa ggagaagaac ttttcactgg agttgtccca attcttgttg
     4201 aattagatgg tgatgttaat gggcacaaat tttctgtccg tggagagggt gaaggtgatg
     4261 ctacaaacgg aaaactcacc cttaaattta tttgcactac tggaaaacta cctgttccgt
     4321 ggccaacact tgtcactact ctgacctatg gtgttcaatg cttttcccgt tatccggatc
     4381 acatgaaacg gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaacgca
     4441 ctatatcttt caaagatgac gggacctaca agacgcgtgc tgaagtcaag tttgaaggtg
     4501 atacccttgt taatcgtatc gagttaaagg gtattgattt taaagaagat ggaaacattc
     4561 ttggacacaa actcgagtac aactttaact cacacaatgt atacatcacg gcagacaaac
     4621 aaaagaatgg aatcaaagct aacttcaaaa ttcgccacaa cgttgaagat ggttccgttc
     4681 aactagcaga ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag
     4741 acaaccatta cctgtcgaca caatctgtcc tttcgaaaga tcccaacgaa aagcgtgacc
     4801 acatggtcct tcttgagttt gtaactgctg ctgggattac acatggcatg gatgagctct
     4861 acaaatgatt aattaaaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     4921 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     4981 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     5041 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     5101 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     5161 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     5221 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     5281 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     5341 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     5401 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     5461 caggatccac tagagcggcc gcctagagct cgctatcagc ctcgactgtg ccttctagtt
     5521 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     5581 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     5641 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     5701 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     5761 cgactagagc ttgcggaacc cttaagcttg tcattcgaat ttagacaccc acatggtggc
     5821 tcatgaccat ctatctataa atggatctga ttttcatgtc tggcagtgta cagaagctaa
     5881 ctgaagaaag gtggaagacc cacaagagtt caaggtaccc aattcgccct atagtgagtc
     5941 gtattacaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     6001 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     6061 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc aaattgtaag
     6121 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     6181 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     6241 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     6301 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     6361 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     6421 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     6481 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     6541 gcttaatgcg ccgctacagg gcgcgtcag
//