LOCUS       sequence-251064-        8106 bp    DNA     circular SYN 11-DEC-2025
DEFINITION  Transposon vector containing an OsTIR IRES Neo cassette driven by 
            the Ef1a promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8106)
  AUTHORS   Strom AR, Biggs RJ, Banigan EJ, Wang X, Chiu K, Herman C, Collado J,
            Yue F, Ritland Politz JC, Tait LJ, Scalzo D, Telling A, Groudine M, 
            Brangwynne CP, Marko JF, Stephens AD
  TITLE     HP1alpha is a chromatin crosslinker that controls nuclear and 
            mitotic chromosome mechanics.
  JOURNAL   Elife. 2021 Jun 9;10. pii: 63972. doi: 10.7554/eLife.63972.
  PUBMED    34106828
REFERENCE   2  (bases 1 to 8106)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 11, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2021-06-9"; volume: "10. pii: 63972. doi"; pages: " 
            10.7554/eLife.63972"
COMMENT     Sequence Label: pEf1a OsTIR ires NEO pA T2BH
FEATURES             Location/Qualifiers
     source          1..8106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1182
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          231..1173
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1130..1150
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1195..2922
                     /codon_start=1
                     /product="rice transport inhibitor response 1 protein"
                     /label=OsTIR1
                     /note="F-box protein that mediates auxin-dependent 
                     interaction of a ubiquitin ligase with an auxin-inducible 
                     degron (Tan et al., 2007)"
                     /translation="MTYFPEEVVEHIFSFLPAQRDRNTVSLVCKVWYEIERLSRRGVFV
                     GNCYAVRAGRVAARFPNVRALTVKGKPHFADFNLVPPDWGGYAGPWIEAAARGCHGLEE
                     LRMKRMVVSDESLELLARSFPRFRALVLISCEGFSTDGLAAVASHCKLLRELDLQENEV
                     EDRGPRWLSCFPDSCTSLVSLNFACIKGEVNAGSLERLVSRSPNLRSLRLNRSVSVDTL
                     AKILLRTPNLEDLGTGNLTDDFQTESYFKLTSALEKCKMLRSLSGFWDASPVCLSFIYP
                     LCAQLTGLNLSYAPTLDASDLTKMISRCVKLQRLWVLDCISDKGLQVVASSCKDLQELR
                     VFPSDFYVAGYSAVTEEGLVAVSLGCPKLNSLLYFCHQMTNAALVTVAKNCPNFTRFRL
                     CILEPGKPDVVTSQPLDEGFGAIVRECKGLQRLSISGLLTDKVFMYIGKYAKQLEMLSI
                     AFAGDSDKGMMHVMNGCKNLRKLEIRDSPFGDAALLGNFARYETMRSLWMSSCNVTLKG
                     CQVLASKMPMLNVEVINERDGSNEMEENHGDLPKVEKLYVYRTTAGARDDAPNFVKIL"
     primer_bind     complement(3150..3167)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3377..3396
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3558..4358
                     /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="MMGIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3618..3637)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4228..4247
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4394..4411)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4400..4607
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4954..4974)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4954..4972)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4993..5009)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4993..5009)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        complement(join(5041..5047,5048..5065,5066..5071))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5086..5107
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5224..5241)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5395..5983)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5475..5494)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6154..6945,6946..7014))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAAKLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6777..6796
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7015..7119)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7146..7601
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7233..7252)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7443..7464
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7727..7749
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7741..7758
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7742..7758
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7768..7787
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7768..7786
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
       61 gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
      121 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
      181 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg
      241 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta
      301 cttccacgcc cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg
      361 tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc
      421 tggcctgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct
      481 gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc
      541 tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg
      601 gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg
      661 cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg
      721 cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca
      781 ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg
      841 aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt
      901 ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc
      961 gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg
     1021 atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg
     1081 taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag
     1141 acagtggttc aaagtttttt tcttccattt caggtgtcgt gactcgacgc caccatgacg
     1201 tacttcccgg aggaggtggt ggagcacatc ttcagcttcc tgccggcgca gcgcgaccgc
     1261 aacacggtct cgctcgtctg caaggtgtgg tacgagatcg agaggctgag ccgccgcggc
     1321 gtcttcgtgg gcaactgcta cgccgtgcgc gccggccgcg tcgccgcgcg gttccccaac
     1381 gtgcgggcgc tcacggtgaa ggggaagccc cacttcgccg acttcaacct cgtgcccccc
     1441 gactggggcg gctacgcggg gccgtggatc gaggcggccg cgaggggatg ccacggcctg
     1501 gaggagctca ggatgaagcg gatggtggtg tccgacgaga gcctcgagct gctggctcgc
     1561 tcgttcccgc ggttcagggc tcttgttctt atcagctgcg aggggttcag cactgacggg
     1621 ctagccgccg tcgcgagcca ttgcaagctt ctgagggagt tggatttgca ggaaaatgaa
     1681 gtggaggatc gagggcctag gtggctttcc tgcttccctg attcctgcac atcacttgtc
     1741 tcattgaatt ttgcctgcat caaaggggag gttaatgctg gttcactgga gagacttgtt
     1801 agcaggtccc caaacctgcg gagtttgagg ctgaatcgat ctgtatcggt agatacactt
     1861 gcaaagatac tactgcgtac ccctaacttg gaggatttgg ggacagggaa tttgacagat
     1921 gacttccaaa ctgagtccta ctttaagctt accagtgctc tggagaaatg caagatgttg
     1981 aggagtttgt ctggattctg ggatgcttct cctgtttgcc tgtcatttat ctaccccctg
     2041 tgtgctcaac tgacaggatt gaacttgagc tatgcaccca cacttgatgc ttctgacctt
     2101 acaaaaatga ttagccgctg tgtgaagctc caacgccttt gggtactgga ttgtatctcg
     2161 gacaaaggct tgcaagtggt ggcctccagt tgcaaagact tgcaagaact cagggtattt
     2221 ccatcagatt tctacgtagc tggttattct gcagtgacag aggagggact tgttgcagta
     2281 tccttgggct gtccaaaact gaactcacta ctgtacttct gtcaccaaat gactaatgct
     2341 gcactagtta ctgtcgccaa gaactgtcca aatttcacac gattcagact ttgtattctt
     2401 gagccaggga agcctgatgt tgtgacaagc caaccattag atgaaggctt tggagctatt
     2461 gttcgtgagt gcaagggatt acaacgtttg tcaatatctg gtcttctcac agacaaagtt
     2521 ttcatgtata ttgggaaata tgcaaaacaa cttgagatgc tttctatagc atttgctggt
     2581 gacagtgata agggtatgat gcatgttatg aatggatgca agaatttaag gaaactggag
     2641 ataagagata gcccgtttgg tgatgctgca ctcttgggga attttgctag gtacgagaca
     2701 atgcgatccc tttggatgtc atcttgcaat gtcacgttaa aggggtgcca agtccttgcg
     2761 tcaaagatgc cgatgctcaa tgttgaggtc ataaatgagc gggatggtag caatgaaatg
     2821 gaggaaaacc atggagatct gcctaaagtg gagaaattat atgtgtaccg cacaactgct
     2881 ggggcgaggg atgatgcacc aaattttgtt aaaatcctat gatctagcaa gtcgacacta
     2941 gcaagtcggg ctgcatctag ggcggccaat tccgcccctc tccccccccc cctccccccc
     3001 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
     3061 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
     3121 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
     3181 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac
     3241 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
     3301 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
     3361 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
     3421 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt
     3481 ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg
     3541 aaaaacacga tgccaccatg atgggaattg aacaagatgg attgcacgca ggttctccgg
     3601 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     3661 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     3721 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     3781 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     3841 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     3901 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     3961 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4021 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4081 ggctcaaggc gcgcatgccc gacggcgacg atctcgtcgt gacccatggc gatgcctgct
     4141 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4201 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4261 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4321 gcatcgcctt ctatcgcctt cttgacgagt tcttctgact cgacaaggcc gcctcggcca
     4381 gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     4441 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     4501 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     4561 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgg aagcttgtga
     4621 aaggctactc gaaatgtttg acccaagtta aacaatttaa aggcaatgct accaaatact
     4681 aattgagtgt atgtaaactt ctgacccact gggaatgtga tgaaagaaat aaaagctgaa
     4741 atgaatcatt ctctctacta ttattctgat atttcacatt cttaaaataa agtggtgatc
     4801 ctaactgacc taagacaggg aatttttact aggattaaat gtcaggaatt gtgaaaaagt
     4861 gagtttaaat gtatttggct aaggtgtatg taaacttccg acttcaactg tatagggatc
     4921 ctctagctag agtcgatcga ccagcttttg ttccctttag tgagggttaa ttgcgcgctt
     4981 ggcgtaatca tggtcatagc tgtttcctgt gtgaaatcgt tatccgctca caattccaca
     5041 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5101 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5161 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5221 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5281 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5341 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5401 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5461 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5521 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaaccgtggc
     5581 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5641 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5701 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagtagcca ctggtaacag
     5761 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5821 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5881 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5941 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6001 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6061 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6121 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6181 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6241 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6301 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6361 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6421 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6481 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6541 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6601 tgtcagaagt aacttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6661 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6721 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6781 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6841 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6901 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6961 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7021 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7081 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7141 acctgacgcg ccctgtagcg gcgcattaag cgcggcggga gtggtggtta cgcgcagcgt
     7201 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     7261 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     7321 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     7381 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     7441 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     7501 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     7561 atttaacgcg aattttaaca aaatattaac gcttacaatt tccattcgcc attcaggctg
     7621 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     7681 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     7741 tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga attggagctc
     7801 ggatccctat acagttgaag tcggaagttt acatacactt aagttggagt cattaaaact
     7861 cgtttttcaa ctactccaca aatttcttgt taacaaacaa tagttttggc aagtcagtta
     7921 ggacatctac tttgtgcatg acacaagtca tttttccaac aattgtttac agacagatta
     7981 tttcacttat aattcactgt atcacaattc cagtgggtca gaagtttaca tacactaagt
     8041 tgactgtgcc tttaaacagc ttggaaaatt ccagaaaatg atgtcatggc tttagaagct
     8101 agatcc
//
