LOCUS       sequence-383244-        6844 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly luciferase promoter expression vector driven only by the 
            promoter element of human lncRNA T-REX17.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6844)
  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 6844)
  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: pGL4.15_pT-REX17
FEATURES             Location/Qualifiers
     source          1..6844
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      980..989
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             986..2635
                     /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             2639..2758
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    2810..2931
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2821..2840)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2875..2894
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3125..3482
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3125..3316
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3148..3168)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3333..3468
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3395..3414
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3513..4550
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4529..4537
                     /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    4574..4622
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4778..4795)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4949..5537)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5029..5048)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5737..6528,6529..6597))
                     /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    6702..6750
                     /note="synthetic polyadenylation signal"
     misc_feature    join(6764..6844,1..11)
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6804..6823
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 aacatttctc tggcctaact ggccggtacc tgagctcgct agcctcgagg atcccaaatc
       61 ccccaaacat tacaactatt tttattttta gttattactt tgcatcattt gtccatatgc
      121 agaaatggtt tcttatagtt ttatattcta cttttttatg cgactttatt ttttgccagc
      181 acagctctca tgttatacta caaaatggct ctaggataga tcacgttgta aaagttgtat
      241 aacactggat attaatattc tattcaggtt tttgctacaa caagcctttt tattagcatg
      301 cttttattct ttttcacttt tctctttggt gggttatatc tttgaaaaaa ttatgaaggc
      361 ttcaatggtt tgtatttaca atgcattcca ctataaaaaa tgagcgtggt ccttcattaa
      421 taaatagtag attcttcacc taaaacaccc agtgatttag catagggatg gattacactc
      481 ttggagaatt tggggtacag acacatttga acgaatgtgt gcttaaaatc cattcatacg
      541 atgtactcat ctcattttga aatggccagg aaaagcagtg gtgtggattt cggcagtgga
      601 gagccctgtg gcaaattccc aacatttcca gagagaaaag ccctcctgga cttactggaa
      661 gagtagagca gaagttaagt catgggggag ggagaacagg aagagactta ggccaaataa
      721 tctgtaacac acacattgtg tgcagcagtt gcaaattcaa agggagctca gtggcccaag
      781 agctaggatg taaagctgac ctctgaatgg gtattatgtc ctatcgaagt ctattgtttc
      841 atgaagccca ggcctaacaa tgtcccattg tggcagatgc aagggcgagt tggttaaatc
      901 ccagttttcc acattttccc ttcatttatc aagatctggc ctcggcggcc aagcttggca
      961 atccggtact gttggtaaag ccaccatgga agatgccaaa aacattaaga agggcccagc
     1021 gccattctac ccactcgaag acgggaccgc cggcgagcag ctgcacaaag ccatgaagcg
     1081 ctacgccctg gtgcccggca ccatcgcctt taccgacgca catatcgagg tggacattac
     1141 ctacgccgag tacttcgaga tgagcgttcg gctggcagaa gctatgaagc gctatgggct
     1201 gaatacaaac catcggatcg tggtgtgcag cgagaatagc ttgcagttct tcatgcccgt
     1261 gttgggtgcc ctgttcatcg gtgtggctgt ggccccagct aacgacatct acaacgagcg
     1321 cgagctgctg aacagcatgg gcatcagcca gcccaccgtc gtattcgtga gcaagaaagg
     1381 gctgcaaaag atcctcaacg tgcaaaagaa gctaccgatc atacaaaaga tcatcatcat
     1441 ggatagcaag accgactacc agggcttcca aagcatgtac accttcgtga cttcccattt
     1501 gccacccggc ttcaacgagt acgacttcgt gcccgagagc ttcgaccggg acaaaaccat
     1561 cgccctgatc atgaacagta gtggcagtac cggattgccc aagggcgtag ccctaccgca
     1621 ccgcaccgct tgtgtccgat tcagtcatgc ccgcgacccc atcttcggca accagatcat
     1681 ccccgacacc gctatcctca gcgtggtgcc atttcaccac ggcttcggca tgttcaccac
     1741 gctgggctac ttgatctgcg gctttcgggt cgtgctcatg taccgcttcg aggaggagct
     1801 attcttgcgc agcttgcaag actataagat tcaatctgcc ctgctggtgc ccacactatt
     1861 tagcttcttc gctaagagca ctctcatcga caagtacgac ctaagcaact tgcacgagat
     1921 cgccagcggc ggggcgccgc tcagcaagga ggtaggtgag gccgtggcca aacgcttcca
     1981 cctaccaggc atccgccagg gctacggcct gacagaaaca accagcgcca ttctgatcac
     2041 ccccgaaggg gacgacaagc ctggcgcagt aggcaaggtg gtgcccttct tcgaggctaa
     2101 ggtggtggac ttggacaccg gtaagacact gggtgtgaac cagcgcggcg agctgtgcgt
     2161 ccgtggcccc atgatcatga gcggctacgt taacaacccc gaggctacaa acgctctcat
     2221 cgacaaggac ggctggctgc acagcggcga catcgcctac tgggacgagg acgagcactt
     2281 cttcatcgtg gaccggctga agagcctgat caaatacaag ggctaccagg tagccccagc
     2341 cgaactggag agcatcctgc tgcaacaccc caacatcttc gacgccgggg tcgccggcct
     2401 gcccgacgac gatgccggcg agctgcccgc cgcagtcgtc gtgctggaac acggtaaaac
     2461 catgaccgag aaggagatcg tggactatgt ggccagccag gttacaaccg ccaagaagct
     2521 gcgcggtggt gttgtgttcg tggacgaggt gcctaaagga ctgaccggca agttggacgc
     2581 ccgcaagatc cgcgagattc tcattaaggc caagaagggc ggcaagatcg ccgtgaattc
     2641 tcacggcttc cctcccgagg tggaggagca ggccgccggc accctgccca tgagctgcgc
     2701 ccaggagagc ggcatggata gacaccctgc tgcttgcgcc agcgccagga tcaacgtcta
     2761 aggccgcgac tctagagtcg gggcggccgg ccgcttcgag cagacatgat aagatacatt
     2821 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     2881 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     2941 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
     3001 taaaacctct acaaatgtgg taaaatcgat aaggatccgt ttgcgtattg ggcgctcttc
     3061 cgctgatctg cgcagcacca tggcctgaaa taacctctga aagaggaact tggttagcta
     3121 ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3181 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3241 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3301 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3361 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3421 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3481 aaaagctcga ttcttctgac actagcgcca ccatgaagaa gcccgaactc accgctacca
     3541 gcgttgaaaa atttctcatc gagaagttcg acagtgtgag cgacctgatg cagttgtcgg
     3601 agggcgaaga gagccgagcc ttcagcttcg atgtcggcgg acgcggctat gtactgcggg
     3661 tgaatagctg cgctgatggc ttctacaaag accgctacgt gtaccgccac ttcgccagcg
     3721 ctgcactacc catccccgaa gtgttggaca tcggcgagtt cagcgagagc ctgacatact
     3781 gcatcagtag acgcgcccaa ggcgttactc tccaagacct ccccgaaaca gagctgcctg
     3841 ctgtgttaca gcctgtcgcc gaagctatgg atgctattgc cgccgccgac ctcagtcaaa
     3901 ccagcggctt cggcccattc gggccccaag gcatcggcca gtacacaacc tggcgggatt
     3961 tcatttgcgc cattgctgat ccccatgtct accactggca gaccgtgatg gacgacaccg
     4021 tgtccgccag cgtagctcaa gccctggacg aactgatgct gtgggccgaa gactgtcccg
     4081 aggtgcgcca cctcgtccat gccgacttcg gcagcaacaa cgtcctgacc gacaacggcc
     4141 gcatcaccgc cgtaatcgac tggtccgaag ctatgttcgg ggacagtcag tacgaggtgg
     4201 ccaacatctt cttctggcgg ccctggctgg cttgcatgga gcagcagact cgctacttcg
     4261 agcgccggca tcccgagctg gccggcagcc ctcgtctgcg agcctacatg ctgcgcatcg
     4321 gcctggatca gctctaccag agcctcgtgg acggcaactt cgacgatgct gcctgggctc
     4381 aaggccgctg cgatgccatc gtccgcagcg gggccggcac cgtcggtcgc acacaaatcg
     4441 ctcgccggag cgcagccgta tggaccgacg gctgcgtcga ggtgctggcc gacagcggca
     4501 accgccggcc cagtacacga ccgcgcgcta aggaggtagg tcgagtttaa actctagaac
     4561 cggtcatggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg
     4621 tgttcgaact agatgctgtc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt
     4681 ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca
     4741 actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4801 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4861 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4921 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4981 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5041 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5101 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5161 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5221 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5281 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5341 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5401 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5461 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5521 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5581 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5641 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5701 ctgacagcgg ccgcaaatgc taaaccactg cagtggttac cagtgcttga tcagtgaggc
     5761 accgatctca gcgatctgcc tatttcgttc gtccatagtg gcctgactcc ccgtcgtgta
     5821 gatcactacg attcgtgagg gcttaccatc aggccccagc gcagcaatga tgccgcgaga
     5881 gccgcgttca ccggcccccg atttgtcagc aatgaaccag ccagcaggga gggccgagcg
     5941 aagaagtggt cctgctactt tgtccgcctc catccagtct atgagctgct gtcgtgatgc
     6001 tagagtaaga agttcgccag tgagtagttt ccgaagagtt gtggccattg ctactggcat
     6061 cgtggtatca cgctcgtcgt tcggtatggc ttcgttcaac tctggttccc agcggtcaag
     6121 ccgggtcaca tgatcaccca tattatgaag aaatgcagtc agctccttag ggcctccgat
     6181 cgttgtcaga agtaagttgg ccgcggtgtt gtcgctcatg gtaatggcag cactacacaa
     6241 ttctcttacc gtcatgccat ccgtaagatg cttttccgtg accggcgagt actcaaccaa
     6301 gtcgttttgt gagtagtgta tacggcgacc aagctgctct tgcccggcgt ctatacggga
     6361 caacaccgcg ccacatagca gtactttgaa agtgctcatc atcgggaatc gttcttcggg
     6421 gcggaaagac tcaaggatct tgccgctatt gagatccagt tcgatatagc ccactcttgc
     6481 acccagttga tcttcagcat cttttacttt caccagcgtt tcggggtgtg caaaaacagg
     6541 caagcaaaat gccgcaaaga agggaatgag tgcgacacga aaatgttgga tgctcatact
     6601 cgtccttttt caatattatt gaagcattta tcagggttac tagtacgtct ctcaaggata
     6661 agtaagtaat attaaggtac gggaggtatt ggacaggccg caataaaata tctttatttt
     6721 cattacatct gtgtgttggt tttttgtgtg aatcgatagt actaacatac gctctccatc
     6781 aaaacaaaac gaaacaaaac aaactagcaa aataggctgt ccccagtgca agtgcaggtg
     6841 ccag
//
