LOCUS       sequence-383248-        7280 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly luciferase enhancer expression vector driven by a minimal 
            CMV-promoter and the extended endoderm distal enhancer of human 
            SOX17.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7280)
  AUTHORS   Landshammer A, Bolondi A, Kretzmer H, Much C, Buschow R, Rose A, Wu 
            HJ, Mackowiak SD, Braendl B, Giesselmann P, Tornisiello R, Parsi KM,
            Huey J, Mielke T, Meierhofer D, Maehr R, Hnisz D, Michor F, Rinn JL,
            Meissner A
  TITLE     T-REX17 is a transiently expressed non-coding RNA essential for 
            human endoderm formation.
  JOURNAL   Elife. 2023 Jan 31;12:e83077. doi: 10.7554/eLife.83077.
  PUBMED    36719724
REFERENCE   2  (bases 1 to 7280)
  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: "Elife."; 
            date: "2023-01-31"; volume: "12:e83077. doi"; pages: " 
            10.7554/eLife.83077"
COMMENT     Sequence Label: pGL4.27_eSOX17
FEATURES             Location/Qualifiers
     source          1..7280
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1355..1386
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      1413..1422
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1419..3068
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     CDS             3072..3191
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    3243..3364
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3254..3273)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3308..3327
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3558..3915
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3558..3749
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3581..3601)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3766..3901
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3828..3847
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3946..4983
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4962..4970
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    5007..5055
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5211..5228)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5382..5970)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5462..5481)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6170..6961,6962..7030))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    7135..7183
                     /note="synthetic polyadenylation signal"
     misc_feature    join(7197..7280,1..8)
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     7237..7256
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 atttctctgg cctaactggc cggtacctga gctcgctagc ctcgaggata tttaaagctt
       61 cgtagcatca ggtgtggtgg agagtattgt ctcttgtggt ttactgagag ggtaaaagct
      121 cttgtaattc ttagtgccaa gcaagcagca tgatcagagc tagataacat ccaaaatagg
      181 tctgttaaaa gagaggaagt caccaaaaga agtgggtaac taagatattt agggttagtg
      241 ctatacatac cagtatgttc cccatccccc aaggttttcc ttctctttaa tcacctcaca
      301 ataggaagtg ctggaaggcg accaggcagg tacctgcact gggtcactcc atgtacctgc
      361 acggggtcac tcatccagct ctgggttttg gacacattgt cagacaaggg taggtttgaa
      421 tgccacttgt ccttaatcat gtcatcacac tgatagcctc ctagcagcaa ccatcagtgt
      481 agtaactata tgtaattcag acctcacctg ggaggtgaat ggaccttggg tgataacact
      541 gaacaatgca ggcacccttc acagtctgaa tttcaccaag tgcctttgag gttgcatcag
      601 tctcgggctt cacaggcaga gggagggaca taggcatagt gatgactttg ttcagagaca
      661 ggcattgatt aggtggcccc taacactggg ggtcaatctg tccaggttta gagggttgct
      721 ttgctgtgat gccacctgca gagttgggac aatgcttagg aaagctgaga cctgcaaccc
      781 taaactggac agaacctaca cagataagct ttgggaagat ctttaacctg actcccttca
      841 tacctgcaag aagccactgt aagattagga aatgtaatta catctgataa agtgattgcc
      901 aagacattgt ttgaagagag ggctttatca gccctattgt atgaagatta atagagatct
      961 ggatcttcta gccactggtc ttgcgcctcc aacagaatga aatctgatta tattagaatg
     1021 tttacatagt agctccatct tccttagact ttcaacactt tataatttat tccctttgtt
     1081 acacactagc ctgggagccc acaaataacc tccagtagcg tgggacccat ccccagggct
     1141 cgactggtga tgtattgact ggcttcacat tctccattgc ttgctctgtg ctcctccctg
     1201 aagcatgtgc cttgcaaagc actgacttca ttacattaca attctgtttc tctgcctctc
     1261 tccccactgg actggaagac ccttgaaaga caggcacttg cttattttgt gttcctggaa
     1321 tcaagatctg gcctcggcgg ccaagcttag acactagagg gtatataatg gaagctcgac
     1381 ttccagcttg gcaatccggt actgttggta aagccaccat ggaagatgcc aaaaacatta
     1441 agaagggccc agcgccattc tacccactcg aagacgggac cgccggcgag cagctgcaca
     1501 aagccatgaa gcgctacgcc ctggtgcccg gcaccatcgc ctttaccgac gcacatatcg
     1561 aggtggacat tacctacgcc gagtacttcg agatgagcgt tcggctggca gaagctatga
     1621 agcgctatgg gctgaataca aaccatcgga tcgtggtgtg cagcgagaat agcttgcagt
     1681 tcttcatgcc cgtgttgggt gccctgttca tcggtgtggc tgtggcccca gctaacgaca
     1741 tctacaacga gcgcgagctg ctgaacagca tgggcatcag ccagcccacc gtcgtattcg
     1801 tgagcaagaa agggctgcaa aagatcctca acgtgcaaaa gaagctaccg atcatacaaa
     1861 agatcatcat catggatagc aagaccgact accagggctt ccaaagcatg tacaccttcg
     1921 tgacttccca tttgccaccc ggcttcaacg agtacgactt cgtgcccgag agcttcgacc
     1981 gggacaaaac catcgccctg atcatgaaca gtagtggcag taccggattg cccaagggcg
     2041 tagccctacc gcaccgcacc gcttgtgtcc gattcagtca tgcccgcgac cccatcttcg
     2101 gcaaccagat catccccgac accgctatcc tcagcgtggt gccatttcac cacggcttcg
     2161 gcatgttcac cacgctgggc tacttgatct gcggctttcg ggtcgtgctc atgtaccgct
     2221 tcgaggagga gctattcttg cgcagcttgc aagactataa gattcaatct gccctgctgg
     2281 tgcccacact atttagcttc ttcgctaaga gcactctcat cgacaagtac gacctaagca
     2341 acttgcacga gatcgccagc ggcggggcgc cgctcagcaa ggaggtaggt gaggccgtgg
     2401 ccaaacgctt ccacctacca ggcatccgcc agggctacgg cctgacagaa acaaccagcg
     2461 ccattctgat cacccccgaa ggggacgaca agcctggcgc agtaggcaag gtggtgccct
     2521 tcttcgaggc taaggtggtg gacttggaca ccggtaagac actgggtgtg aaccagcgcg
     2581 gcgagctgtg cgtccgtggc cccatgatca tgagcggcta cgttaacaac cccgaggcta
     2641 caaacgctct catcgacaag gacggctggc tgcacagcgg cgacatcgcc tactgggacg
     2701 aggacgagca cttcttcatc gtggaccggc tgaagagcct gatcaaatac aagggctacc
     2761 aggtagcccc agccgaactg gagagcatcc tgctgcaaca ccccaacatc ttcgacgccg
     2821 gggtcgccgg cctgcccgac gacgatgccg gcgagctgcc cgccgcagtc gtcgtgctgg
     2881 aacacggtaa aaccatgacc gagaaggaga tcgtggacta tgtggccagc caggttacaa
     2941 ccgccaagaa gctgcgcggt ggtgttgtgt tcgtggacga ggtgcctaaa ggactgaccg
     3001 gcaagttgga cgcccgcaag atccgcgaga ttctcattaa ggccaagaag ggcggcaaga
     3061 tcgccgtgaa ttctcacggc ttccctcccg aggtggagga gcaggccgcc ggcaccctgc
     3121 ccatgagctg cgcccaggag agcggcatgg atagacaccc tgctgcttgc gccagcgcca
     3181 ggatcaacgt ctaaggccgc gactctagag tcggggcggc cggccgcttc gagcagacat
     3241 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     3301 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     3361 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     3421 tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgtttgcgta
     3481 ttgggcgctc ttccgctgat ctgcgcagca ccatggcctg aaataacctc tgaaagagga
     3541 acttggttag ctaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     3601 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3661 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3721 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3781 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3841 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3901 ctaggctttt gcaaaaagct cgattcttct gacactagcg ccaccatgaa gaagcccgaa
     3961 ctcaccgcta ccagcgttga aaaatttctc atcgagaagt tcgacagtgt gagcgacctg
     4021 atgcagttgt cggagggcga agagagccga gccttcagct tcgatgtcgg cggacgcggc
     4081 tatgtactgc gggtgaatag ctgcgctgat ggcttctaca aagaccgcta cgtgtaccgc
     4141 cacttcgcca gcgctgcact acccatcccc gaagtgttgg acatcggcga gttcagcgag
     4201 agcctgacat actgcatcag tagacgcgcc caaggcgtta ctctccaaga cctccccgaa
     4261 acagagctgc ctgctgtgtt acagcctgtc gccgaagcta tggatgctat tgccgccgcc
     4321 gacctcagtc aaaccagcgg cttcggccca ttcgggcccc aaggcatcgg ccagtacaca
     4381 acctggcggg atttcatttg cgccattgct gatccccatg tctaccactg gcagaccgtg
     4441 atggacgaca ccgtgtccgc cagcgtagct caagccctgg acgaactgat gctgtgggcc
     4501 gaagactgtc ccgaggtgcg ccacctcgtc catgccgact tcggcagcaa caacgtcctg
     4561 accgacaacg gccgcatcac cgccgtaatc gactggtccg aagctatgtt cggggacagt
     4621 cagtacgagg tggccaacat cttcttctgg cggccctggc tggcttgcat ggagcagcag
     4681 actcgctact tcgagcgccg gcatcccgag ctggccggca gccctcgtct gcgagcctac
     4741 atgctgcgca tcggcctgga tcagctctac cagagcctcg tggacggcaa cttcgacgat
     4801 gctgcctggg ctcaaggccg ctgcgatgcc atcgtccgca gcggggccgg caccgtcggt
     4861 cgcacacaaa tcgctcgccg gagcgcagcc gtatggaccg acggctgcgt cgaggtgctg
     4921 gccgacagcg gcaaccgccg gcccagtaca cgaccgcgcg ctaaggaggt aggtcgagtt
     4981 taaactctag aaccggtcat ggccgcaata aaatatcttt attttcatta catctgtgtg
     5041 ttggtttttt gtgtgttcga actagatgct gtcgaccgat gcccttgaga gccttcaacc
     5101 cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct
     5161 tctttatcat gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact
     5221 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5281 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5341 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5401 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5461 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5521 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5581 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5641 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5701 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5761 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5821 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5881 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5941 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6001 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6061 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6121 gagtaaactt ggtctgacag cggccgcaaa tgctaaacca ctgcagtggt taccagtgct
     6181 tgatcagtga ggcaccgatc tcagcgatct gcctatttcg ttcgtccata gtggcctgac
     6241 tccccgtcgt gtagatcact acgattcgtg agggcttacc atcaggcccc agcgcagcaa
     6301 tgatgccgcg agagccgcgt tcaccggccc ccgatttgtc agcaatgaac cagccagcag
     6361 ggagggccga gcgaagaagt ggtcctgcta ctttgtccgc ctccatccag tctatgagct
     6421 gctgtcgtga tgctagagta agaagttcgc cagtgagtag tttccgaaga gttgtggcca
     6481 ttgctactgg catcgtggta tcacgctcgt cgttcggtat ggcttcgttc aactctggtt
     6541 cccagcggtc aagccgggtc acatgatcac ccatattatg aagaaatgca gtcagctcct
     6601 tagggcctcc gatcgttgtc agaagtaagt tggccgcggt gttgtcgctc atggtaatgg
     6661 cagcactaca caattctctt accgtcatgc catccgtaag atgcttttcc gtgaccggcg
     6721 agtactcaac caagtcgttt tgtgagtagt gtatacggcg accaagctgc tcttgcccgg
     6781 cgtctatacg ggacaacacc gcgccacata gcagtacttt gaaagtgctc atcatcggga
     6841 atcgttcttc ggggcggaaa gactcaagga tcttgccgct attgagatcc agttcgatat
     6901 agcccactct tgcacccagt tgatcttcag catcttttac tttcaccagc gtttcggggt
     6961 gtgcaaaaac aggcaagcaa aatgccgcaa agaagggaat gagtgcgaca cgaaaatgtt
     7021 ggatgctcat actcgtcctt tttcaatatt attgaagcat ttatcagggt tactagtacg
     7081 tctctcaagg ataagtaagt aatattaagg tacgggaggt attggacagg ccgcaataaa
     7141 atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agtactaaca
     7201 tacgctctcc atcaaaacaa aacgaaacaa aacaaactag caaaataggc tgtccccagt
     7261 gcaagtgcag gtgccagaac
//
