LOCUS       sequence-92270-e        7668 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses eGFP from a 400 bp human FOXA2 promoter to select floor 
            plate and ventral midbrain progenitor cells during development .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7668)
  AUTHORS   Aguila JC, Blak A, van Arensbergen J, Sousa A, Vazquez N, Aduriz A, 
            Gayosso M, Paz Lopez Mato M, Lopez de Maturana R, Hedlund E, Sonntag
            K, Sanchez Pernaute R
  TITLE     Selection Based on FOXA2 Expression Is Not Sufficient to Enrich for 
            Dopamine Neurons From Human Pluripotent Stem Cells.
  JOURNAL   Stem Cells Transl Med. 2014 Jul 14. pii: sctm.2014-0011.
  PUBMED    25024431
REFERENCE   2  (bases 1 to 7668)
  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: "Stem Cells 
            Transl Med. 2014 Jul 14. pii: sctm.2014-0011."
COMMENT     Sequence Label: Lenti.pFOXA2.GFP
FEATURES             Location/Qualifiers
     source          1..7668
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..131
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(132..200,201..992)
                     /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(350..369)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1163..1751
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1652..1671
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1905..1922
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2039..2060
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2075..2080,2081..2098,2099..2105)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2113..2129
                     /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     2118..2140
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2137..2153
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2137..2153
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2172..2192
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2174..2192
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     LTR             2399..2656
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2700..2825
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3318..3551
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3735..3779
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    4046..4163
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     regulatory      4591..4600
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4597..4599,4600..4602,4603..5316)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4597..4599,4600..4602,4603..5313)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4642..4663)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4903..4922)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5250..5271
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5332..5921
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5385..5405)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5803..5814)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6009..6642
                     /label=5' LTR
                     /note="5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    6714..6835
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6751..6770)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6805..6824
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6854..6989
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7021..7040)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7022..7040)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7050..7067)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7050..7066)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7059..7081)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7211..7666
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7298..7317)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7508..7529
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
       61 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
      121 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
      181 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
      241 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
      301 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
      361 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
      421 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
      481 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
      541 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
      601 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
      661 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
      721 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
      781 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
      841 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
      901 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
      961 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     1021 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     1081 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     1141 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     1201 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     1261 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     1321 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     1381 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     1441 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     1501 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     1561 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     1621 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     1681 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     1741 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     1801 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     1861 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     1921 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     1981 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     2041 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     2101 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     2161 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttaat
     2221 gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt agcaacatgc
     2281 cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg
     2341 tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca ctgaattgga
     2401 ggcgtggcct gggcgggact ggggagtggc gagccctcag atcctgcata taagcagctg
     2461 ctttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     2521 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     2581 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     2641 tggaaaatct ctagcagtgg cgcccgaaca gggacctgaa agcgaaaggg aaaccagagc
     2701 tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac
     2761 tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga
     2821 gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag
     2881 ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat
     2941 tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc
     3001 tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa
     3061 ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga
     3121 tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga
     3181 cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta
     3241 aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa
     3301 aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact
     3361 atgggcgcag cctcaatgac gctgacggta caggccagac aattattgtc tggtatagtg
     3421 cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca
     3481 gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat
     3541 caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct
     3601 tggaatgcta gttggagtaa taaatctctg gaacagattg gaatcacacg acctggatgg
     3661 agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc
     3721 aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt
     3781 ggaattggtt taacataaca aattggctgt ggtatataaa attattcata atgatagtag
     3841 gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc
     3901 agggatattc accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc
     3961 ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga
     4021 acggatctcg acggtatcgg ttaactttta aaagaaaagg ggggattggg gggtacagtg
     4081 caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac
     4141 aaattacaaa aattcaaaat tttatcgatc acgagactag cctcgagcaa ggtccagcca
     4201 ggttttcaga accaagaaag agcctccaca tccaaacacc tgcaatatcc ccccactcca
     4261 aatctgggct cacaggctaa cccagaacag aagacaattt ttgaacccaa gagctgctgg
     4321 ggaaataaaa gtatacgatt gctggagttt ctaatttcta ttaagcagtc cctctggaag
     4381 acagagagga cagagacgct cttgaagtca actccatatg ccccatcatt gattcctgga
     4441 ttcttctctc ctcacccctc cctccccatc tcctgccctg tttgttttag ttacgaaatg
     4501 ctgtgggcac ctcggttgtg actgaaaagt aaccttgaaa cacgccggcc tgaatatcag
     4561 aggcaaatct cagcctccca accaccggtc gccaccatgg tgagcaaggg cgaggagctg
     4621 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     4681 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     4741 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     4801 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     4861 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     4921 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     4981 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     5041 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     5101 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     5161 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     5221 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     5281 gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgcgtcga caatcaacct
     5341 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     5401 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     5461 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     5521 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     5581 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     5641 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     5701 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     5761 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     5821 gaccttcctt cccgcggcct gctgcccggc tctgcggcct cttccgcgtc ttcgccttcg
     5881 ccctcagacg agtcggatct ccctttgggc cgcctccccg ccntggaatt cgagctcggt
     5941 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     6001 ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat
     6061 ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc cagggatcag
     6121 atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag agaaggtaga
     6181 agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc atgggatgga
     6241 tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat
     6301 ggcccgagag ctgcatccgg agtacttcaa gaactgctga catcgagctt gctacaaggg
     6361 actttccgct ggggactttc cagggaggcg tggcctgggc gggactgggg agtggcgagc
     6421 cctcagatgc tgcatataag cagctgcttt ttgcttgtac tgggtctctc tggttagacc
     6481 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     6541 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     6601 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc
     6661 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt
     6721 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     6781 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     6841 tctggctcta gctatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     6901 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     6961 gaagtagtga ggaggctttt ttggaggcct aggcttttgc gtcgagacgt acccaattcg
     7021 ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg
     7081 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     7141 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     7201 gaatggcgcg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     7261 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     7321 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     7381 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     7441 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     7501 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     7561 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     7621 caaaaattta acgcgaattt taacaaaata ttaacgttta caatttcc
//
