LOCUS       Exported                8634 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  sfGFP and iCreERT2 fused by unskippable linker.
ACCESSION   .
VERSION     .
KEYWORDS    pCAG-sfGFP-iCre-ERT2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8634)
  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 8634)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8634
                     /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..3064
                     /label=chicken beta-actin promoter
     intron          3065..4081
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     4005..4027
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     4089..4108
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             4150..4863
                     /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(4180..4208)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4803..4823
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             5992..6927
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     primer_bind     complement(6991..7011)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7409..7420)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7565..7582)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7571..7795
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7992..8011)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7993..8011)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8018..8035)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8018..8034)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8027..8049)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(8176..8631)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8313..8334)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8525..8544
                     /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 ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg
     2941 gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa
     3001 gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg
     3061 ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg
     3121 cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc
     3181 tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg
     3241 tgaaagcctt gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggggtgc
     3301 gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga
     3361 gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc
     3421 cgggggcggt gccccgcggt gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg
     3481 tgtgtgcgtg ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct
     3541 gcacccccct ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg
     3601 gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg
     3661 ggcggggccg cctcgggccg gggagggctc gggggagggg cgcggcggcc cccggagcgc
     3721 cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag
     3781 ggcgcaggga cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg
     3841 caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg
     3901 ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg
     3961 tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg
     4021 tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca
     4081 gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattgctagc
     4141 gccgccacca tgagcaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa
     4201 ttagatggtg atgttaatgg gcacaaattt tctgtccgtg gagagggtga aggtgatgct
     4261 acaaacggaa aactcaccct taaatttatt tgcactactg gaaaactacc tgttccgtgg
     4321 ccaacacttg tcactactct gacctatggt gttcaatgct tttcccgtta tccggatcac
     4381 atgaaacggc atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaacgcact
     4441 atatctttca aagatgacgg gacctacaag acgcgtgctg aagtcaagtt tgaaggtgat
     4501 acccttgtta atcgtatcga gttaaagggt attgatttta aagaagatgg aaacattctt
     4561 ggacacaaac tcgagtacaa ctttaactca cacaatgtat acatcacggc agacaaacaa
     4621 aagaatggaa tcaaagctaa cttcaaaatt cgccacaacg ttgaagatgg ttccgttcaa
     4681 ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac
     4741 aaccattacc tgtcgacaca atctgtcctt tcgaaagatc ccaacgaaaa gcgtgaccac
     4801 atggtccttc ttgagtttgt aactgctgct gggattacac atggcatgga tgagctctac
     4861 aaaggaggat ctggcggccg ctctgccaca aacttcagcc tgctgaagca ggccggcgac
     4921 gccgctgaaa atcctggccc tgctcctgga agcaccaacc tgctgactgt gcaccaaaac
     4981 ctgcctgccc tccctgtgga tgccacctct gatgaagtca ggaagaacct gatggacatg
     5041 ttcagggaca ggcaggcctt ctctgaacac acctggaaga tgctcctgtc tgtgtgcaga
     5101 tcctgggctg cctggtgcaa gctgaacaac aggaaatggt tccctgctga acctgaggat
     5161 gtgagggact acctcctgta cctgcaagcc agaggcctgg ctgtgaagac catccaacag
     5221 cacctgggcc agctcaacat gctgcacagg agatctggcc tgcctcgccc ttctgactcc
     5281 aatgctgtgt ccctggtgat gaggagaatc agaaaggaga atgtggatgc tggggagaga
     5341 gccaagcagg ccctggcctt tgaacgcact gactttgacc aagtcagatc cctgatggag
     5401 aactctgaca gatgccagga catcaggaac ctggccttcc tgggcattgc ctacaacacc
     5461 ctgctgcgca ttgccgaaat tgccagaatc agagtgaagg acatctcccg caccgatggt
     5521 gggagaatgc tgatccacat tggcaggacc aagaccctgg tgtccacagc tggtgtggag
     5581 aaggccctgt ccctgggggt taccaagctg gtggagagat ggatctctgt gtctggtgtg
     5641 gctgatgacc ccaacaacta cctgttctgc cgggtcagaa agaatggtgt ggctgcccct
     5701 tctgccacct cccaactgtc cacccgggcc ctggaaggga tctttgaggc cacccaccgc
     5761 ctgatctatg gtgccaagga tgactctggg cagagatacc tggcctggtc tggccactct
     5821 gccagagtgg gtgctgccag ggacatggcc agggctggtg tgtccatccc tgaaatcatg
     5881 caggctggtg gctggaccaa tgtgaacatt gtgatgaact acatcagaaa cctggactct
     5941 gagactgggg ccatggtgag gctgctagag gatggggacc tcgagccatc tgctggagac
     6001 atgagagctg ccaacctttg gccaagcccg ctcatgatca aacgctctaa gaagaacagc
     6061 ctggccttgt ccctgacggc cgaccagatg gtcagtgcct tgttggatgc tgagcccccc
     6121 atactctatt ccgagtatga tcctaccaga cccttcagtg aagcttcgat gatgggctta
     6181 ctgaccaacc tggcagacag ggagctggtt cacatgatca actgggcgaa gagggtgcca
     6241 ggctttgtgg atttgaccct ccatgatcag gtccaccttc tagaatgtgc ctggctagag
     6301 atcctgatga ttggtctcgt ctggcgctcc atggagcacc cagtgaagct actgtttgct
     6361 cctaacttgc tcttggacag gaaccaggga aaatgtgtag agggcatggt ggagatcttc
     6421 gacatgctgc tggctacatc atctcggttc cgcatgatga atctgcaggg agaggagttt
     6481 gtgtgcctca aatctattat tttgcttaat tctggagtgt acacatttct gtccagcacc
     6541 ctgaagtctc tggaagagaa ggaccatatc caccgagtcc tggacaagat cacagacact
     6601 ttgatccacc tgatggccaa ggcaggcctg accctgcagc agcagcacca gcggctggcc
     6661 cagctcctcc tcatcctctc ccacatcagg cacatgagta acaaaggcat ggagcatctg
     6721 tacagcatga agtgcaagaa cgtggtgccc ctctatgacc tgctgctgga ggcggcggac
     6781 gcccaccgcc tacatgcgcc cactagccgt ggaggggcat ccgtggagga gacggaccaa
     6841 agccacttgg ccactgcggg ctctacttca tcgcattcct tgcaaaagta ttacatcacg
     6901 ggggaggcag agggtttccc tgccacagct tgattaatta aaacctctgg attacaaaat
     6961 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     7021 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     7081 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     7141 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     7201 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     7261 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     7321 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     7381 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     7441 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     7501 gatctccctt tgggccgcct ccccgcagga tccactagag cggccgccta gagctcgcta
     7561 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     7621 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     7681 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     7741 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     7801 gaggcggaaa gaaccagctg gggctcgact agagcttgcg gaacccttaa gcttgtcatt
     7861 cgaatttaga cacccacatg gtggctcatg accatctatc tataaatgga tctgattttc
     7921 atgtctggca gtgtacagaa gctaactgaa gaaaggtgga agacccacaa gagttcaagg
     7981 tacccaattc gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc
     8041 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     8101 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     8161 tgaatggcga atggcaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     8221 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     8281 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     8341 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     8401 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     8461 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     8521 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     8581 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcag
//
