LOCUS       sequence-148975-        5341 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  positive control for lncRNA expression assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5341)
  TITLE     pTREtight reporter plasmids--Roderic Guigo
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5341)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pTre-Tight-luciferase-Hottip (522)
FEATURES             Location/Qualifiers
     source          1..5341
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        4..318
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    12..30
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    48..66
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    83..101
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    119..137
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    155..173
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    190..208
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    226..244
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     291..315
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     350..369
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        350..368
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(873..890)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1087..1168
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1179..1196)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1350..1938)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1430..1449)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2109..2900,2901..2969))
                     /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     2732..2751
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2970..3074)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3091..3287
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3138..3273
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3323..3332
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3329..4981
                     /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"
     polyA_signal    5025..5146
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5036..5055)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5310..5328
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagttta ctccctatca gtgatagaga acgtatgtcg agtttactcc ctatcagtga
       61 tagagaacga tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttactc
      121 cctatcagtg atagagaacg tatgtcgagt ttactcccta tcagtgatag agaacgtatg
      181 tcgagtttat ccctatcagt gatagagaac gtatgtcgag tttactccct atcagtgata
      241 gagaacgtat gtcgaggtag gcgtgtacgg tgggaggcct atataagcag agctcgttta
      301 gtgaaccgtc agatcgcctg gagaattcga gctcggtacc cggggatcct aatacgactc
      361 actataggga gacccaagct ggctagcgtt taaacttaag cttggcaccg agctcggatc
      421 ccgcctttat cccagtgggg cccagacccg cgcaaccagg cggggagggg aggtgggcgc
      481 gcgattgggt tgcgatctgg agcagtgggg acaggtcagg aaccggcgcg tatttctgca
      541 gtgagaccac aggacggaca tcggcgcctt cggcttcgat ggagttgcga ttttgctctt
      601 tccagggaaa cagtggcagg gtgtttgctg cttatcggtt cctgcggata tgcctgggtc
      661 ccaggacatt ccactggagg cttggactgc atttaggagc ccctatccct tccctgtcca
      721 cactgttagt gagcaatttc atatgtttgc atttagaccg atagactcag gacgactcat
      781 cacacacaca cacagtgtac actgacacac tcacattcgc acacttaggt atacagcctg
      841 atccttgctc tgacctggta acaacgcttc ctcctcgact gtgccttcta gaattcgagc
      901 tcggtacccg gggatcctct agtcagctga cgcgtgctag cgcggccgca tcgataagct
      961 tgtcgacgat atctctagag gatcataatc agccatacca catttgtaga ggttttactt
     1021 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     1081 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     1141 ttcacaaata aagcattttt ttcactgcct cgagcttcct cgctcactga ctcgctgcgc
     1201 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     1261 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     1321 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1381 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     1441 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     1501 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     1561 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     1621 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     1681 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     1741 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     1801 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     1861 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     1921 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     1981 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     2041 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     2101 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     2161 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     2221 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     2281 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     2341 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     2401 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     2461 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     2521 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     2581 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     2641 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     2701 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     2761 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     2821 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     2881 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     2941 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     3001 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     3061 ataggggttc cgcgcacatt tcccagatct tgcatctcaa ttagtcagca accatagtcc
     3121 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3181 atcgctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3241 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag cttaagcttg
     3301 gcaatccggt actgttggta aagccaccat ggaagatgcc aaaaacatta agaagggccc
     3361 agcgccattc tacccactcg aagacgggac cgccggcgag cagctgcaca aagccatgaa
     3421 gcgctacgcc ctggtgcccg gcaccatcgc ctttaccgac gcacatatcg aggtggacat
     3481 tacctacgcc gagtacttcg agatgagcgt tcggctggca gaagctatga agcgctatgg
     3541 gctgaataca aaccatcgga tcgtggtgtg cagcgagaat agcttgcagt tcttcatgcc
     3601 cgtgttgggt gccctgttca tcggtgtggc tgtggcccca gctaacgaca tctacaacga
     3661 gcgcgagctg ctgaacagca tgggcatcag ccagcccacc gtcgtattcg tgagcaagaa
     3721 agggctgcaa aagatcctca acgtgcaaaa gaagctaccg atcatacaaa agatcatcat
     3781 catggatagc aagaccgact accagggctt ccaaagcatg tacaccttcg tgacttccca
     3841 tttgccaccc ggcttcaacg agtacgactt cgtgcccgag agcttcgacc gggacaaaac
     3901 catcgccctg atcatgaaca gtagtggcag taccggattg cccaagggcg tagccctacc
     3961 gcaccgcacc gcttgtgtcc gattcagtca tgcccgcgac cccatcttcg gcaaccagat
     4021 catccccgac accgctatcc tcagcgtggt gccatttcac cacggcttcg gcatgttcac
     4081 cacgctgggc tacttgatct gcggctttcg ggtcgtgctc atgtaccgct tcgaggagga
     4141 gctattcttg cgcagcttgc aagactataa gattcaatct gccctgctgg tgcccacact
     4201 atttagcttc ttcgctaaga gcactctcat cgacaagtac gacctaagca acttgcacga
     4261 gatcgccagc ggcggggcgc cgctcagcaa ggaggtaggt gaggccgtgg ccaaacgctt
     4321 ccacctacca ggcatccgcc agggctacgg cctgacagaa acaaccagcg ccattctgat
     4381 cacccccgaa ggggacgaca agcctggcgc agtaggcaag gtggtgccct tcttcgaggc
     4441 taaggtggtg gacttggaca ccggtaagac actgggtgtg aaccagcgcg gcgagctgtg
     4501 cgtccgtggc cccatgatca tgagcggcta cgttaacaac cccgaggcta caaacgctct
     4561 catcgacaag gacggctggc tgcacagcgg cgacatcgcc tactgggacg aggacgagca
     4621 cttcttcatc gtggaccggc tgaagagcct gatcaaatac aagggctacc aggtagcccc
     4681 agccgaactg gagagcatcc tgctgcaaca ccccaacatc ttcgacgccg gggtcgccgg
     4741 cctgcccgac gacgatgccg gcgagctgcc cgccgcagtc gtcgtgctgg aacacggtaa
     4801 aaccatgacc gagaaggaga tcgtggacta tgtggccagc caggttacaa ccgccaagaa
     4861 gctgcgcggt ggtgttgtgt tcgtggacga ggtgcctaaa ggactgaccg gcaagttgga
     4921 cgcccgcaag atccgcgaga ttctcattaa ggccaagaag ggcggcaaga tcgccgtgta
     4981 ataattctag agtcggggcg gccggccgct tcgagcagac atgataagat acattgatga
     5041 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     5101 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     5161 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     5221 cctctacaaa tgtggtaaaa tcgataagga tccgaaaagt gccacctgac gtctaagaaa
     5281 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc
     5341 a
//
