LOCUS       sequence-442720-        6457 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 6457)
  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 6457)
  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_v2
FEATURES             Location/Qualifiers
     source          1..6457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        33..532
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(68..91)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     465..484
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             572..1171
                     /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     1727..1743
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1727..1743
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2736..2752)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2737..2753)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             complement(2765..3364)
                     /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(3365..3576)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     primer_bind     complement(3595..3614)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3596..3614)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3624..3641)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3624..3640)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3633..3655)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3782..4237
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3869..3888)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4079..4100
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4263..4367
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4368..4436,4437..5228)
                     /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(4586..4605)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5399..5987
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5888..5907
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6141..6158
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6275..6296
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6311..6316,6317..6334,6335..6341)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6349..6365
                     /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     6354..6376
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6373..6389
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6373..6389
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6408..6428
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6410..6428
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 cggccgctat tcgtcgacct cgaaattcta ccgggtaggg gaggcgcttt tcccaaggca
       61 gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg
      121 gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc
      181 cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc
      241 gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag
      301 caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc
      361 cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg
      421 gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca
      481 aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacctgcat
      541 ccatctagat ctcgagcagc tgaagcttac catgaccgag tacaagccca cggtgcgcct
      601 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
      661 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
      721 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
      781 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
      841 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
      901 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
      961 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     1021 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     1081 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     1141 ctggtgcatg acccgcaagc ccggtgcctg acgcccgccc cacgacccgc agcgcccgac
     1201 cgaaaggagc gcacgacccc atgcatcgat gatatcagat ccccgggatg cagaaattga
     1261 tgatctatta aacaataaag atgtccacta aaatggaagt ttttcctgtc atactttgtt
     1321 aagaagggtg agaacagagt acctacattt tgaatggaag gattggagct acgggggtgg
     1381 gggtggggtg ggattagata aatgcctgct ctttactgaa ggctctttac tattgcttta
     1441 tgataatgtt tcatagttgg atatcataat ttaaacaagc aaaaccaaat taagggccag
     1501 ctcattcctc ccactcatga tctatagatc tatagatctc tcgtgggatc attgtttttc
     1561 tcttgattcc cactttgtgg ttctaagtac tgtggtttcc aaatgtgtca gtttcatagc
     1621 ctgaagaacg agatcagcag cctctgttcc acatacactt cattctcagt attgttttgc
     1681 caagttctaa ttccatcaga agctggtcga ctctagatgc ggccgctcta gaactagtgg
     1741 atccgtatac catgaattcg atccctaacc ctaaccctaa ccctaaccct aaccctaacc
     1801 ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct aaccctaacc
     1861 ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct aaccctaacc
     1921 ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct aaccctaacc
     1981 ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct aaccctaacc
     2041 ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct aaccctaacc
     2101 ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct aaccctaacc
     2161 ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct aaccctaacc
     2221 ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct aaccctaacc
     2281 ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct aaccctaacc
     2341 ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct aaccctaacc
     2401 ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct aaccctaacc
     2461 ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct aaccctaacc
     2521 ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct aaccctaacc
     2581 ctaaccctaa ccctaaccct aaccctaacc ttaaccctaa ccctaaccct aaccctaacc
     2641 ctaaccctaa ccctaaccct aaccctaacc ctaaccctaa ccctaaccct aaccctaacc
     2701 ctaaccctaa ccctaaccct aaccatcaag cttatcgata ccgtcgacct cgaggggggg
     2761 cccgtcaggc accgggcttg cgggtcatgc accaggtgcg cggtccttcg ggcacctcga
     2821 cgtcggcggt gacggtgaag ccgagccgct cgtagaaggg gaggttgcgg ggcgcggagg
     2881 tctccaggaa ggcgggcacc ccggcgcgct cggccgcctc cactccgggg agcacgacgg
     2941 cgctgcccag acccttgccc tggtggtcgg gcgagacgcc gacggtggcc aggaaccacg
     3001 cgggctcctt gggccggtgc ggcgccagga ggccttccat ctgttgctgc gcggccagcc
     3061 gggaaccgct caactcggcc atgcgcgggc cgatctcggc gaacaccgcc cccgcttcga
     3121 cgctctccgg cgtggtccag accgccaccg cggcgccgtc gtccgcgacc cacaccttgc
     3181 cgatgtcgag cccgacgcgc gtgaggaaga gttcttgcag ctcggtgacc cgctcgatgt
     3241 ggcggtccgg gtcgacggtg tggcgcgtgg cggggtagtc ggcgaacgcg gcggcgaggg
     3301 tgcgtacggc ccgggggacg tcgtcgcggg tggcgaggcg caccgtgggc ttgtactcgg
     3361 tcatctgtgt tctggcggca aacccgttgc gaaaaagaac gttcacggcg actactgcac
     3421 ttatatacgg ttctccccca ccctcgggaa aaaggcggag ccagtacacg acatcacttt
     3481 cccagtttac cccgcgccac cttctctagg caccggttca attgccgacc cctcccccca
     3541 acttctcggg gactgtgggc gatgtgcgct ctgcccggta ccgtacccaa ttcgccctat
     3601 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     3661 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     3721 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     3781 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3841 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3901 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3961 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     4021 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4081 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4141 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4201 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
     4261 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4321 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     4381 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4441 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     4501 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4561 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4621 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4681 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4741 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     4801 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4861 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     4921 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     4981 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5041 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5101 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5161 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5221 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     5281 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5341 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5401 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5461 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5521 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5581 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5641 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5701 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5761 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5821 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5881 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5941 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6001 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6061 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     6121 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     6181 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     6241 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     6301 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     6361 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
     6421 actaaaggga acaaaagctg gagctccacc gcggtgg
//
