LOCUS       sequence-383996-        6458 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ~100 repeats of the human telomere seed sequence fused to the 
            puromycin resistance gene; digest with BstZ17I-HF and KpnI-HF to 
            obtain transfectable construct for chromosomal arm loss induction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6458)
  AUTHORS   Girish V, Lakhani AA, Thompson SL, Scaduto CM, Brown LM, Hagenson 
            RA, Sausville EL, Mendelson BE, Kandikuppa PK, Lukow DA, Yuan ML, 
            Stevens EC, Lee SN, Schukken KM, Akalu SM, Vasudevan A, Zou C, 
            Salovska B, Li W, Smith JC, Taylor AM, Martienssen RA, Liu Y, Sun R,
            Sheltzer JM
  TITLE     Oncogene-like addiction to aneuploidy in human cancers.
  JOURNAL   Science. 2023 Jul 6:eadg4521. doi: 10.1126/science.adg4521.
  PUBMED    37410869
REFERENCE   2  (bases 1 to 6458)
  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: "Science. 
            2023 Jul 6:eadg4521. doi: 10.1126/science.adg4521."
COMMENT     Sequence Label: EF1a_Puro_Telo_v1
FEATURES             Location/Qualifiers
     source          1..6458
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     33..53
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        35..53
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        115..614
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(150..173)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     547..566
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             654..1253
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     1809..1825
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1809..1825
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2819..2835)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2820..2836)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             complement(2848..3447)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     promoter        complement(3448..3659)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     primer_bind     complement(3678..3697)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3679..3697)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3707..3724)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3707..3723)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3716..3738)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3865..4320
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3952..3971)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4162..4183
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4346..4450
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4451..4519,4520..5311)
                     /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     complement(4669..4688)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5482..6070
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5971..5990
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6224..6241
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6358..6379
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6394..6399,6400..6417,6418..6424)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6432..6448
                     /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)."
     primer_bind     join(6437..6458,1)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6456..6458,1..14)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6456..6458,1..14)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa
       61 agctggagct ccaccgcggt ggcggccgct attcgtcgac ctcgaaattc taccgggtag
      121 gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt
      181 ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac
      241 cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt
      301 tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca
      361 ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca
      421 gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt
      481 ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag
      541 gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat
      601 ctccgggcct ttcgacctgc atccatctag atctcgagca gctgaagctt accatgaccg
      661 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
      721 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
      781 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
      841 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
      901 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
      961 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     1021 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     1081 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     1141 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     1201 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc
     1261 cccacgaccc gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg atgatatcag
     1321 atccccggga tgcagaaatt gatgatctat taaacaataa agatgtccac taaaatggaa
     1381 gtttttcctg tcatactttg ttaagaaggg tgagaacaga gtacctacat tttgaatgga
     1441 aggattggag ctacgggggt gggggtgggg tgggattaga taaatgcctg ctctttactg
     1501 aaggctcttt actattgctt tatgataatg tttcatagtt ggatatcata atttaaacaa
     1561 gcaaaaccaa attaagggcc agctcattcc tcccactcat gatctataga tctatagatc
     1621 tctcgtggga tcattgtttt tctcttgatt cccactttgt ggttctaagt actgtggttt
     1681 ccaaatgtgt cagtttcata gcctgaagaa cgagatcagc agcctctgtt ccacatacac
     1741 ttcattctca gtattgtttt gccaagttct aattccatca gaagctggtc gactctagat
     1801 gcggccgctc tagaactagt ggatccgtat accatgaatt cgatccctaa ccctaaccct
     1861 aaccctaacc ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct
     1921 aaccctaacc ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct
     1981 aaccctaacc ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct
     2041 aaccctaacc ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa cactaaccct
     2101 aaccctaacc ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct
     2161 aaccctaacc ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct
     2221 aaccctaacc ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct
     2281 aaccctaacc ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct
     2341 aaccctaacc ctaaccctaa ccctaaccct aaccctaacc mtaaccctaa ccctaaccct
     2401 aaccctaacc ctaaccctaa ccctaaccct acaccctaac cctaacccta accctaaccc
     2461 taaccctaac cctaacccta accctaaccc taaccctaac cctaacccta accctaaccc
     2521 taaccctaac cctaacccta accctaaccc taaccctaac cctaacccta accctaaccc
     2581 taaccctaac cctaacccta accctaaccc taaccctaac cctaacccta accctaaccc
     2641 taaccctaac cctaacccta accctaaccc taaccctaac cctaacccta accytaaccc
     2701 taaccctaac cctaacccta accctaaccc taaccctaac cctaacccta accctaaccc
     2761 taaccctaac cctaacccta accctaaccc taaccctaac cctaaccatc aagcttatcg
     2821 ataccgtcga cctcgagggg gggcccgtca ggcaccgggc ttgcgggtca tgcaccaggt
     2881 gcgcggtcct tcgggcacct cgacgtcggc ggtgacggtg aagccgagcc gctcgtagaa
     2941 ggggaggttg cggggcgcgg aggtctccag gaaggcgggc accccggcgc gctcggccgc
     3001 ctccactccg gggagcacga cggcgctgcc cagacccttg ccctggtggt cgggcgagac
     3061 gccgacggtg gccaggaacc acgcgggctc cttgggccgg tgcggcgcca ggaggccttc
     3121 catctgttgc tgcgcggcca gccgggaacc gctcaactcg gccatgcgcg ggccgatctc
     3181 ggcgaacacc gcccccgctt cgacgctctc cggcgtggtc cagaccgcca ccgcggcgcc
     3241 gtcgtccgcg acccacacct tgccgatgtc gagcccgacg cgcgtgagga agagttcttg
     3301 cagctcggtg acccgctcga tgtggcggtc cgggtcgacg gtgtggcgcg tggcggggta
     3361 gtcggcgaac gcggcggcga gggtgcgtac ggcccggggg acgtcgtcgc gggtggcgag
     3421 gcgcaccgtg ggcttgtact cggtcatctg tgttctggcg gcaaacccgt tgcgaaaaag
     3481 aacgttcacg gcgactactg cacttatata cggttctccc ccaccctcgg gaaaaaggcg
     3541 gagccagtac acgacatcac tttcccagtt taccccgcgc caccttctct aggcaccggt
     3601 tcaattgccg acccctcccc ccaacttctc ggggactgtg ggcgatgtgc gctctgcccg
     3661 gtaccgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt
     3721 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     3781 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     3841 tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3901 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3961 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     4021 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     4081 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     4141 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     4201 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     4261 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     4321 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4381 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4441 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4501 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4561 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4621 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4681 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4741 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     4801 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4861 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     4921 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     4981 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5041 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5101 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5161 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5221 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5281 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     5341 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5401 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5461 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5521 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5581 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5641 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5701 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5761 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5821 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5881 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5941 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6001 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6061 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6121 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     6181 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     6241 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     6301 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     6361 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     6421 gttgtgtgga attgtgagcg gataacaatt tcacacag
//
