LOCUS       sequence-393095-        7439 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of full length mTRF2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7439)
  AUTHORS   Kwan JZJ, Nguyen TF, Uzozie AC, Budzynski MA, Cui J, Lee JMC, Van 
            Petegem F, Lange PF, Teves SS
  TITLE     RNA Polymerase II transcription independent of TBP in murine 
            embryonic stem cells.
  JOURNAL   Elife. 2023 Mar 30;12:e83810. doi: 10.7554/eLife.83810.
  PUBMED    36995326
REFERENCE   2  (bases 1 to 7439)
  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: "2023-03-30"; volume: "12:e83810. doi"; pages: " 
            10.7554/eLife.83810"
COMMENT     Sequence Label: TRF2-HA
FEATURES             Location/Qualifiers
     source          1..7439
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     172..194
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     186..203
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     187..203
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   333..395
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        848..1059
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             1072..1340
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             1926..1952
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    1971..2543
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2138..2155)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2365..2384
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2552..3346
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLAATGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2606..2625)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3216..3235
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3406..3426)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3468..3589
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3505..3524)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3559..3578
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4352..4386
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(4877..4893)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4877..4893)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4890..4912)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4901..4917
                     /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)."
     promoter        complement(join(4925..4931,4932..4949,4950..4955))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4970..4991
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5108..5125)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5279..5867)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5359..5378)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6038..6829,6830..6898))
                     /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     6661..6680
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6899..7003)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(7029..7439,1..45))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7166..7187)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7378..7397
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca
       61 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
      121 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
      181 gacgttgtaa aacgacggcc agtgagcgcg cctcgttcat tcacgttttt gaacccgtgg
      241 aggacgggca gactcgcggt gcaaatgtgt tttacagcgt gatggagcag atgaagatgc
      301 tcgacacgct gcagaacacg cagctagatt aaccctagaa agataatcat attgtgacgt
      361 acgttaaaga taatcatgtg taaaattgac gcatgtgttt tatcggtctg tatatcgagg
      421 tttatttatt aatttgaata gatattaagt tttattatat ttacacttac atactaataa
      481 taaattcaac aaacaattta tttatgttta tttatttatt aaaaaaaaca aaaactcaaa
      541 atttcttcta taaagtaaca aaacttttat gagggacagc ccccccccaa agcccccagg
      601 gatgtaatta cgtccctccc ccgctagggg gcagcagcga gccgcccggg gctccgctcc
      661 ggtccggcgc tccccccgca tccccgagcc ggcagcgtgc ggggacagcc cgggcacggg
      721 gaaggtggca cgggatcgct ttcctctgaa cgcttctcgc tgctctttga gcctgcagac
      781 acctgggggg atacggggaa aaggcctcca cggccaagga tctgcgatcg ctccggtgcc
      841 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
      901 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
      961 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
     1021 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc
     1081 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc
     1141 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta
     1201 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc
     1261 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct
     1321 gttctgcgcc gttacagatc caagctgtga ccggcgccta ctctagaatg gatgcagaca
     1381 gtgatgttgc attggacatt ttaattacaa atgtagtctg tgtttttaga acaagatgcc
     1441 atttgaactt aaggaagatt gctttggagg gagcaaatgt aatttataag cgtgatgttg
     1501 ggaaagtatt aatgaagctt agaaaaccta gaattacagc tacaatttgg tcctcaggaa
     1561 aaattatttg cactggagca acaagtgaag aagaagctaa atttggtgcc aggcgtttag
     1621 cccgtagtct gcagaaacta ggttttcagg tcatcttcac agattttaag gtggtgaatg
     1681 ttttggcagt gtgtaacatg ccctttgaga tccgtttgcc agaatttaca aagaacaata
     1741 gacctcatgc cagctatgaa cctgaacttc atcctgccgt gtgctatcgg ataaagtctc
     1801 taagagctac attacagata ttttcaacag gaagcatcac agtgacaggg cccaatgtaa
     1861 aggctgtggc gactgccgtg gaacagatct acccgttcgt gtttgaaagc aggaaggaga
     1921 ttttataccc atacgatgtt ccagattacg cttaaggatc cgcggccgcg cccctctccc
     1981 tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc
     2041 tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc
     2101 cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt
     2161 ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct
     2221 gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa
     2281 aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt
     2341 tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag
     2401 gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt
     2461 acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt
     2521 ttcctttgaa aaacacgata atacctccgg aatgattgaa caagatggat tgcacgcagg
     2581 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     2641 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     2701 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     2761 ggccgcgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     2821 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     2881 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     2941 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     3001 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     3061 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     3121 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     3181 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     3241 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     3301 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagtcg acaatcaacc
     3361 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     3421 gctatgtgga tacgctgctt taatgccttt gtatcatgcg ttaactaaac ttgtttattg
     3481 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3541 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     3601 attgactcaa atgatgtcaa ttagtctatc agaagctcat ctggtctccc ttccggggga
     3661 caagacatcc ctgtttaata tttaaacagc agtgttccca aactgggttc ttatatccct
     3721 tgctctggtc aaccaggttg cagggtttcc tgtcctcaca ggaacgaagt ccctaaagaa
     3781 acagtggcag ccaggtttag ccccggaatt gactggattc cttttttagg gcccattggt
     3841 atggcttttt ccccgtatcc ccccaggtgt ctgcaggctc aaagagcagc gagaagcgtt
     3901 cagaggaaag cgatcccgtg ccaccttccc cgtgcccggg ctgtccccgc acgctgccgg
     3961 ctcggggatg cggggggagc gccggaccgg agcggagccc cgggcggctc gctgctgccc
     4021 cctagcgggg gagggacgta attacatccc tgggggcttt gggggggggc tgtccctgat
     4081 atctataaca agaaaatata tatataataa gttatcacgt aagtagaaca tgaaataaca
     4141 atataattat cgtatgagtt aaatcttaaa agtcacgtaa aagataatca tgcgtcattt
     4201 tgactcacgc ggtcgttata gttcaaaatc agtgacactt accgcattga caagcacgcc
     4261 tcacgggagc tccaagcggc gactgagatg tcctaaatgc acagcgacgg attcgcgcta
     4321 tttagaaaga gagagcaata tttcaagaat gcatgcgtca attttacgca gactatcttt
     4381 ctagggttaa tctagctgca tcaggatcat atcgtcgggt cttttttccg gctcagtcat
     4441 cgcccaagct ggcgctatct gggcatcggg gaggaagaag cccgtgcctt ttcccgcgag
     4501 gttgaagcgg catggaaaga gtttgccgag gatgactgct gctgcattga cgttgagcga
     4561 aaacgcacgt ttaccatgat gattcgggaa ggtgtggcca tgcacgcctt taacggtgaa
     4621 ctgttcgttc aggccacctg ggataccagt tcgtcgcggc ttttccggac acagttccgg
     4681 atggtcagcc cgaagcgcat cagcaacccg aacaataccg gcgacagccg gaactgccgt
     4741 gccggtgtgc agattaatga cagcggtgcg gcgctgggat attacgtcag cgaggacggg
     4801 tatcctggct ggatgccgca gaaatggaca tggatacccc gtgagttacc cggcgggcgc
     4861 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     4921 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4981 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5041 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5101 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5161 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5221 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5281 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5341 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5401 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5461 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5521 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5581 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5641 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5701 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5761 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5821 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5881 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5941 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6001 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6061 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6121 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6181 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6241 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6301 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6361 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6421 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6481 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6541 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6601 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6661 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6721 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6781 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6841 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6901 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6961 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7021 tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     7081 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     7141 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     7201 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     7261 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     7321 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     7381 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcg
//
