LOCUS       sequence-383246-        6645 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly luciferase enhancer expression vector driven by a minimal 
            CMV-promoter and the core endoderm distal enhancer of human SOX17.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6645)
  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 6645)
  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_eSOX17.2
FEATURES             Location/Qualifiers
     source          1..6645
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     31..50
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        794..825
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      852..861
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             858..2507
                     /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             2511..2630
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    2682..2803
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2693..2712)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2747..2766
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2997..3354
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2997..3188
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3020..3040)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3205..3340
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3267..3286
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3385..4422
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4401..4409
                     /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    4446..4494
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4650..4667)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4821..5409)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4901..4920)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5609..6400,6401..6469))
                     /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    6574..6622
                     /note="synthetic polyadenylation signal"
     misc_feature    join(6636..6645,1..82)
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
ORIGIN
        1 ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt
       61 gcaggtgcca gaacatttct ctggcctaac tggccggtac ctgagctcgc tagcctcgag
      121 gatactgggg gtcaatctgt ccaggtttag agggttgctt tgctgtgatg ccacctgcag
      181 agttgggaca atgcttagga aagctgagac ctgcaaccct aaactggaca gaacctacac
      241 agataagctt tgggaagatc tttaacctga ctcccttcat acctgcaaga agccactgta
      301 agattaggaa atgtaattac atctgataaa gtgattgcca agacattgtt tgaagagagg
      361 gctttatcag ccctattgta tgaagattaa tagagatctg gatcttctag ccactggtct
      421 tgcgcctcca acagaatgaa atctgattat attagaatgt ttacatagta gctccatctt
      481 ccttagactt tcaacacttt ataatttatt ccctttgtta cacactagcc tgggagccca
      541 caaataacct ccagtagcgt gggacccatc cccagggctc gactggtgat gtattgactg
      601 gcttcacatt ctccattgct tgctctgtgc tcctccctga agcatgtgcc ttgcaaagca
      661 ctgacttcat tacattacaa ttctgtttct ctgcctctct ccccactgga ctggaagacc
      721 cttgaaagac aggcacttgc ttattttgtg ttcctggaat caagatctgg cctcggcggc
      781 caagcttaga cactagaggg tatataatgg aagctcgact tccagcttgg caatccggta
      841 ctgttggtaa agccaccatg gaagatgcca aaaacattaa gaagggccca gcgccattct
      901 acccactcga agacgggacc gccggcgagc agctgcacaa agccatgaag cgctacgccc
      961 tggtgcccgg caccatcgcc tttaccgacg cacatatcga ggtggacatt acctacgccg
     1021 agtacttcga gatgagcgtt cggctggcag aagctatgaa gcgctatggg ctgaatacaa
     1081 accatcggat cgtggtgtgc agcgagaata gcttgcagtt cttcatgccc gtgttgggtg
     1141 ccctgttcat cggtgtggct gtggccccag ctaacgacat ctacaacgag cgcgagctgc
     1201 tgaacagcat gggcatcagc cagcccaccg tcgtattcgt gagcaagaaa gggctgcaaa
     1261 agatcctcaa cgtgcaaaag aagctaccga tcatacaaaa gatcatcatc atggatagca
     1321 agaccgacta ccagggcttc caaagcatgt acaccttcgt gacttcccat ttgccacccg
     1381 gcttcaacga gtacgacttc gtgcccgaga gcttcgaccg ggacaaaacc atcgccctga
     1441 tcatgaacag tagtggcagt accggattgc ccaagggcgt agccctaccg caccgcaccg
     1501 cttgtgtccg attcagtcat gcccgcgacc ccatcttcgg caaccagatc atccccgaca
     1561 ccgctatcct cagcgtggtg ccatttcacc acggcttcgg catgttcacc acgctgggct
     1621 acttgatctg cggctttcgg gtcgtgctca tgtaccgctt cgaggaggag ctattcttgc
     1681 gcagcttgca agactataag attcaatctg ccctgctggt gcccacacta tttagcttct
     1741 tcgctaagag cactctcatc gacaagtacg acctaagcaa cttgcacgag atcgccagcg
     1801 gcggggcgcc gctcagcaag gaggtaggtg aggccgtggc caaacgcttc cacctaccag
     1861 gcatccgcca gggctacggc ctgacagaaa caaccagcgc cattctgatc acccccgaag
     1921 gggacgacaa gcctggcgca gtaggcaagg tggtgccctt cttcgaggct aaggtggtgg
     1981 acttggacac cggtaagaca ctgggtgtga accagcgcgg cgagctgtgc gtccgtggcc
     2041 ccatgatcat gagcggctac gttaacaacc ccgaggctac aaacgctctc atcgacaagg
     2101 acggctggct gcacagcggc gacatcgcct actgggacga ggacgagcac ttcttcatcg
     2161 tggaccggct gaagagcctg atcaaataca agggctacca ggtagcccca gccgaactgg
     2221 agagcatcct gctgcaacac cccaacatct tcgacgccgg ggtcgccggc ctgcccgacg
     2281 acgatgccgg cgagctgccc gccgcagtcg tcgtgctgga acacggtaaa accatgaccg
     2341 agaaggagat cgtggactat gtggccagcc aggttacaac cgccaagaag ctgcgcggtg
     2401 gtgttgtgtt cgtggacgag gtgcctaaag gactgaccgg caagttggac gcccgcaaga
     2461 tccgcgagat tctcattaag gccaagaagg gcggcaagat cgccgtgaat tctcacggct
     2521 tccctcccga ggtggaggag caggccgccg gcaccctgcc catgagctgc gcccaggaga
     2581 gcggcatgga tagacaccct gctgcttgcg ccagcgccag gatcaacgtc taaggccgcg
     2641 actctagagt cggggcggcc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     2701 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     2761 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     2821 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     2881 ctacaaatgt ggtaaaatcg ataaggatcc gtttgcgtat tgggcgctct tccgctgatc
     2941 tgcgcagcac catggcctga aataacctct gaaagaggaa cttggttagc taccttctga
     3001 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3061 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3121 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3181 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3241 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     3301 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     3361 gattcttctg acactagcgc caccatgaag aagcccgaac tcaccgctac cagcgttgaa
     3421 aaatttctca tcgagaagtt cgacagtgtg agcgacctga tgcagttgtc ggagggcgaa
     3481 gagagccgag ccttcagctt cgatgtcggc ggacgcggct atgtactgcg ggtgaatagc
     3541 tgcgctgatg gcttctacaa agaccgctac gtgtaccgcc acttcgccag cgctgcacta
     3601 cccatccccg aagtgttgga catcggcgag ttcagcgaga gcctgacata ctgcatcagt
     3661 agacgcgccc aaggcgttac tctccaagac ctccccgaaa cagagctgcc tgctgtgtta
     3721 cagcctgtcg ccgaagctat ggatgctatt gccgccgccg acctcagtca aaccagcggc
     3781 ttcggcccat tcgggcccca aggcatcggc cagtacacaa cctggcggga tttcatttgc
     3841 gccattgctg atccccatgt ctaccactgg cagaccgtga tggacgacac cgtgtccgcc
     3901 agcgtagctc aagccctgga cgaactgatg ctgtgggccg aagactgtcc cgaggtgcgc
     3961 cacctcgtcc atgccgactt cggcagcaac aacgtcctga ccgacaacgg ccgcatcacc
     4021 gccgtaatcg actggtccga agctatgttc ggggacagtc agtacgaggt ggccaacatc
     4081 ttcttctggc ggccctggct ggcttgcatg gagcagcaga ctcgctactt cgagcgccgg
     4141 catcccgagc tggccggcag ccctcgtctg cgagcctaca tgctgcgcat cggcctggat
     4201 cagctctacc agagcctcgt ggacggcaac ttcgacgatg ctgcctgggc tcaaggccgc
     4261 tgcgatgcca tcgtccgcag cggggccggc accgtcggtc gcacacaaat cgctcgccgg
     4321 agcgcagccg tatggaccga cggctgcgtc gaggtgctgg ccgacagcgg caaccgccgg
     4381 cccagtacac gaccgcgcgc taaggaggta ggtcgagttt aaactctaga accggtcatg
     4441 gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgttcgaa
     4501 ctagatgctg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg
     4561 cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag
     4621 gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4681 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4741 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4801 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4861 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4921 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4981 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5041 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5101 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5161 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5221 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5281 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5341 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5401 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5461 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5521 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagc
     5581 ggccgcaaat gctaaaccac tgcagtggtt accagtgctt gatcagtgag gcaccgatct
     5641 cagcgatctg cctatttcgt tcgtccatag tggcctgact ccccgtcgtg tagatcacta
     5701 cgattcgtga gggcttacca tcaggcccca gcgcagcaat gatgccgcga gagccgcgtt
     5761 caccggcccc cgatttgtca gcaatgaacc agccagcagg gagggccgag cgaagaagtg
     5821 gtcctgctac tttgtccgcc tccatccagt ctatgagctg ctgtcgtgat gctagagtaa
     5881 gaagttcgcc agtgagtagt ttccgaagag ttgtggccat tgctactggc atcgtggtat
     5941 cacgctcgtc gttcggtatg gcttcgttca actctggttc ccagcggtca agccgggtca
     6001 catgatcacc catattatga agaaatgcag tcagctcctt agggcctccg atcgttgtca
     6061 gaagtaagtt ggccgcggtg ttgtcgctca tggtaatggc agcactacac aattctctta
     6121 ccgtcatgcc atccgtaaga tgcttttccg tgaccggcga gtactcaacc aagtcgtttt
     6181 gtgagtagtg tatacggcga ccaagctgct cttgcccggc gtctatacgg gacaacaccg
     6241 cgccacatag cagtactttg aaagtgctca tcatcgggaa tcgttcttcg gggcggaaag
     6301 actcaaggat cttgccgcta ttgagatcca gttcgatata gcccactctt gcacccagtt
     6361 gatcttcagc atcttttact ttcaccagcg tttcggggtg tgcaaaaaca ggcaagcaaa
     6421 atgccgcaaa gaagggaatg agtgcgacac gaaaatgttg gatgctcata ctcgtccttt
     6481 ttcaatatta ttgaagcatt tatcagggtt actagtacgt ctctcaagga taagtaagta
     6541 atattaaggt acgggaggta ttggacaggc cgcaataaaa tatctttatt ttcattacat
     6601 ctgtgtgttg gttttttgtg tgaatcgata gtactaacat acgct
//
