LOCUS Exported 8374 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION expresses eGFP from an about 1350 bp human FOXA2 promoter to select floor plate and ventral midbrain progenitor cells during development. ACCESSION . VERSION . KEYWORDS Lenti.pFOXA2.5'.GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8374) 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 8374) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8374 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 27..131 /gene="bla" /label=AmpR promoter CDS 132..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 2075..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" gap 4917..4945 /estimated_length=29 regulatory 5297..5306 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 5303..6022 /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 5303..6019 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(5348..5369) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(5609..5628) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 5956..5977 /label=EGFP-C /note="EGFP, forward primer" misc_feature 6038..6627 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(6091..6111) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6509..6520) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 6715..7348 /label=5' LTR /note="5' long terminal repeat (LTR) from HIV-1" polyA_signal 7420..7541 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(7457..7476) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 7511..7530 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 7560..7695 /label=SV40 ori /note="SV40 origin of replication" primer_bind 7622..7641 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(7727..7746) /label=T7 /note="T7 promoter, forward primer" promoter complement(7728..7746) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(7756..7773) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(7756..7772) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7765..7787) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 7917..8372 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(8004..8023) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 8214..8235 /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 cctccccacc tcctgccctg tttgttttag ttacgaaatg 4501 ctgtgggcac ctcggttgtg actgaaaagt aaccttgaaa cacgccggcc tgaatatcag 4561 aggcaaatct cagcctccca accgtcggcc gctgctagag gggctgcttg cgccaggcgc 4621 cggccgcccc actgcgggtc cctggcggcc ggtgtctgag gagtcggaga gccgaggcgg 4681 ccagaccgtg cgccccgcgc ttctcccgag gccgttccgg gtctgaactg taacagggag 4741 gggcctcgca ggagcagcag cgggcgagtt aaaggtgtgt acgtagtttt tctaaatagg 4801 acagcgcttt gcaaattggc tgtcatggac ttgctgtttt gacaganaat gagcactgag 4861 ggcgaanaaa cccggcataa ataaatccgg ctttcntagg gntctgccgg gcactgnnnn 4921 nnnnnnnnnn nnnnnnnnnn nnnnntcctc ctccccaccc ccccactagc ccccgcgccg 4981 cgcgcttccc gccgcctcgg ggcagctcgg cactgccaac ctcagccgcc acccggcagc 5041 ccgtgagcgc ggcggcggcg ggtgctccct accgcggggc gctggcctgg gcctcctccg 5101 ctcctcgccc ccccccacct cccgcccact tccaactacc gcctccggcc tgcccaggga 5161 gagagaggga gtggagccca gggagaggga gcgcgagaga gggagggagg aggggacggt 5221 gctttggctg actttttttt aaaagagggt gggggtgggg ggtgattgct ggtcgtttgt 5281 tgtggctgta ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt 5341 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga 5401 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa 5461 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag 5521 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta 5581 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt 5641 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga 5701 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat 5761 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga 5821 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc 5881 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa 5941 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg 6001 catggacgag ctgtacaagt aaagcggccg cgtcgacaat caacctctgg attacaaaat 6061 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc 6121 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt 6181 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg 6241 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg 6301 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc 6361 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt 6421 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct 6481 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg 6541 cggcctgctg cccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc 6601 ggatctccct ttgggccgcc tccccgccnt ggaattcgag ctcggtacct ttaagaccaa 6661 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag 6721 ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag 6781 gctacttccc tgattggcag aactacacac cagggccagg gatcagatat ccactgacct 6841 ttggatggtg ctacaagcta gtaccagttg agcaagagaa ggtagaagaa gccaatgaag 6901 gagagaacac ccgcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag 6961 aagtattaga gtggaggttt gacagccgcc tagcatttca tcacatggcc cgagagctgc 7021 atccggagta cttcaagaac tgctgacatc gagcttgcta caagggactt tccgctgggg 7081 actttccagg gaggcgtggc ctgggcggga ctggggagtg gcgagccctc agatgctgca 7141 tataagcagc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg 7201 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg 7261 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc 7321 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat 7381 ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat 7441 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 7501 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta 7561 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt 7621 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag 7681 gcttttttgg aggcctaggc ttttgcgtcg agacgtaccc aattcgccct atagtgagtc 7741 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt 7801 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga 7861 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcgacgc 7921 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac 7981 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt 8041 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc 8101 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc 8161 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact 8221 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg 8281 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc 8341 gaattttaac aaaatattaa cgtttacaat ttcc //