LOCUS       sequence-364928-        7003 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human POT1 with a zz affinity tag, YBBR labeling site, and
            TEV cleavage site in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7003)
  AUTHORS   Jack A, Kim Y, Strom AR, Lee DSW, Williams B, Schaub JM, Kellogg EH,
            Finkelstein IJ, Ferro LS, Yildiz A, Brangwynne CP
  TITLE     Compartmentalization of telomeres through DNA-scaffolded phase 
            separation.
  JOURNAL   Dev Cell. 2022 Jan 24;57(2):277-290.e9. doi: 
            10.1016/j.devcel.2021.12.017.
  PUBMED    35077681
REFERENCE   2  (bases 1 to 7003)
  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"; doi: 
            "10.1016/j.devcel.2021.12"; journalName: "Dev Cell"; date: 
            "2022-01-24- 24"; volume: "57"; issue: "2"; pages: "277-290.e9"
COMMENT     Sequence Label: pOmniBac zz TEV YBBR POT1
FEATURES             Location/Qualifiers
     source          1..7003
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      319..907
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     808..827
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_recomb     1163..1990
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             1163..1287
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(1325..1930)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     mobile_element  2019..2242
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2309..2842)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3031..3036,3037..3053,3054..3059))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(3130..3163)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3240..3331
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3256..3273
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             3398..3421
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             3464..3637
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAYYEILHLPNLNEGQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             3638..3811
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAEKLNDAQAPK"
     CDS             3869..3889
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3911..3931
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    6000..6134
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6037..6056)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6091..6110
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  6255..6420
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     misc_recomb     join(6427..7003,1..129)
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           6515..6887
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(join(6544..7003,1..129))
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
ORIGIN
        1 tcgtcgtcgt cccaaccctc gtcgttagaa gttgcttccg aagacgattt tgccatagcc
       61 acacgacgcc tattaattgt gtcggctaac acgtccgcga tcaaatttgt agttgagctt
      121 tttggaatta ccggttgact tgggtcaact gtcagaccaa gtttactcat atatacttta
      181 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
      241 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
      301 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
      361 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
      421 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
      481 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      541 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
      601 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      661 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      721 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
      781 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      841 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      901 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
      961 tcacatgttc tttcctgcgt tatcccctga ttgacttggg tcgctcttcc tgtggatgcg
     1021 caggtatgta caggaagagg tttatactaa actgttacat tgcaaacgtg gtttcgtgtg
     1081 ccaagtgtga aaaccgatgt ttaatcaagg ctctgacgca tttctacaac cacgactcta
     1141 agtgtgtggg tgaagtcatg catgcatctt ttaatcaaat cccaagatgt gtataaacca
     1201 ccaaactgcc aaaaaatgaa aactgtcgac aagctctgtc cgtttgctgg caactgcaag
     1261 ggtctcaatc ctatttgtaa ttattgaata ataaaacaat tataaatgtc aaatttgttt
     1321 tttattaacg atacaaacca aacgcaacaa gaacatttgt agtattatct ataattgaaa
     1381 acgcgtagtt ataatcgctg aggtaatatt taaaatcatt ttcaaatgat tcacagttaa
     1441 tttgcgacaa tataatttta ttttcacata aactagacgc cttgtcgtct tcttcttcgt
     1501 attccttctc tttttcattt ttctcttcat aaaaattaac atagttatta tcgtatccat
     1561 atatgtatct atcgtataga gtaaattttt tgttgtcata aatatatatg tcttttttaa
     1621 tggggtgtat agtaccgctg cgcatagttt ttctgtaatt tacaacagtg ctattttctg
     1681 gtagttcttc ggagtgtgtt gctttaatta ttaaatttat ataatcaatg aatttgggat
     1741 cgtcggtttt gtacaatatg ttgccggcat agtacgcagc ttcttctagt tcaattacac
     1801 cattttttag cagcaccgga ttaacataac tttccaaaat gttgtacgaa ccgttaaaca
     1861 aaaacagttc acctcccttt tctatactat tgtctgcgag cagttgtttg ttgttaaaaa
     1921 taacagccat tgtaatgaga cgcacaaact aatatcacaa actggaaatg tctatcaata
     1981 tatagttgct gattgcgcag atgccctgcg taagcgggtg tgggcggaca ataaagtctt
     2041 aaactgaaca aaatagatct aaactatgac aataaagtct taaactagac agaatagttg
     2101 taaactgaaa tcagtccagt tatgctgtga aaaagcatac tggacttttg ttatggctaa
     2161 agcaaactct tcattttctg aagtgcaaat tgcccgtcgt attaaagagg ggcgtggcca
     2221 agggcatgta aagactatat tcgcggcgtt gtgacaattt accgaacaac tccgcggccg
     2281 ggaagccgat ctcggcttga acgaattgtt aggtggcggt acttgggtcg atatcaaagt
     2341 gcatcacttc ttcccgtatg cccaactttg tatagagagc cactgcggga tcgtcaccgt
     2401 aatctgcttg cacgtagatc acataagcac caagcgcgtt ggcctcatgc ttgaggagat
     2461 tgatgagcgc ggtggcaatg ccctgcctcc ggtgctcgcc ggagactgcg agatcataga
     2521 tatagatctc actacgcggc tgctcaaact tgggcagaac gtaagccgcg agagcgccaa
     2581 caaccgcttc ttggtcgaag gcagcaagcg cgatgaatgt cttactacgg agcaagttcc
     2641 cgaggtaatc ggagtccggc tgatgttggg agtaggtggc tacgtctccg aactcacgac
     2701 cgaaaagatc aagagcagcc cgcatggatt tgacttggtc agggccgagc ctacatgtgc
     2761 gaatgatgcc catacttgag ccacctaact ttgttttagg gcgactgccc tgctgcgtaa
     2821 catcgttgct gctgcgtaac atcgttgctg ctccataaca tcaaacatcg acccacggcg
     2881 taacgcgctt gctgcttgga tgcccgaggc atagactgta caaaaaaaca gtcataacaa
     2941 gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt cggtcaaggt tctggaccag
     3001 ttgcgtgagc gcatacgcta cttgcattac agtttacgaa ccgaacaggc ttatgtcaac
     3061 tgggttcgtg ccttcatccg tttccacggt gtgcgtcacc cggcaacctt gggcagcagc
     3121 gaagtcgcca taacttcgta tagcatacat tatacgaagt tatctgtaac tataacggtc
     3181 ctaaggtagc gagtttaaac actagtatcg attcgcgacc tactccggaa tattaataga
     3241 tcatggagat aattaaaatg ataaccatct cgcaaataaa taagtatttt actgttttcg
     3301 taacagtttt gtaataaaaa aacctataaa tattccggat tattcatacc gtcccaccat
     3361 cgggcgcgga tctcggtccg aaaccatgtc gtactaccat caccatcacc atcaccatca
     3421 cggttctatg tgcatggcag gccttgcgca acacgatgaa gccgtggaca acaaattcaa
     3481 caaagaacaa caaaacgcgt actatgagat cttacattta cctaacttaa acgaaggaca
     3541 acgaaacgcc ttcatccaaa gtttaaaaga tgacccaagc caaagcgcta accttttagc
     3601 agaagctaaa aagctaaatg atgctcaggc gccgaaagta gacaacaaat tcaacaaaga
     3661 acaacaaaac gcgttctatg agatcttaca tttacctaac ttaaacgaag aacaacgaaa
     3721 cgccttcatc caaagtttaa aagatgaccc aagccaaagc gctaaccttt tagcagaagc
     3781 tgaaaagcta aatgatgctc aggcgccgaa agtagacgca aattccgcgg ggaagtcaac
     3841 cgattatgat attccaacta ctgctagcga gaacctatat ttccagggag attatgatat
     3901 ccctacgact gagaatcttt attttcaggg catggactca ttggagttta tcgcatcaaa
     3961 gctcgcaatg tctttggttc cagcaacaaa ttatatatat acacccctga atcaacttaa
     4021 gggtggtaca attgtcaatg tctatggtgt tgtgaagttc tttaagcccc catatctaag
     4081 caaaggaact gattattgct cagttgtaac tattgtggac cagacaaatg taaaactaac
     4141 ttgcctgctc tttagtggaa actatgaagc ccttccaata atttataaaa atggagatat
     4201 tgttcgcttt cacaggctga agattcaagt atataaaaag gagactcagg gtatcaccag
     4261 ctctggcttt gcatctttga cgtttgaggg aactttggga gcccctatca tacctcgcac
     4321 ttcaagcaag tattttaact tcactactga ggaccacaaa atggtagaag ccttacgtgt
     4381 ttgggcatct actcatatgt caccgtcttg gacattacta aaattgtgtg atgttcagcc
     4441 aatgcagtat tttgacctga cttgtcagct cttgggcaaa gcagaagtgg acggagcatc
     4501 atttcttcta aaggtatggg atggcaccag gacaccattt ccatcttgga gagtcttaat
     4561 acaagacctt gttcttgaag gtgatttaag tcacatccat cggctacaaa atctgacaat
     4621 agacatttta gtctacgata accatgttca tgtggcaaga tctctgaagg ttggaagctt
     4681 tcttagaatc tatagccttc ataccaaact tcaatcaatg aattcagaga atcagacaat
     4741 gttaagttta gagtttcatc ttcatggagg taccagttac ggtcggggaa tcagggtctt
     4801 gccagaaagt aactctgatg tggatcaact gaaaaaggat ttagaatctg caaatttgac
     4861 agccaatcag cattcagatg ttatctgtca atcagaacct gacgacagct ttccaagctc
     4921 tggatcagta tcattatacg aggtagaaag atgtcaacag ctatctgcta caatacttac
     4981 agatcatcag tatttggaga ggacaccact atgtgccatt ttgaaacaaa aagctcctca
     5041 acaataccgc atccgagcaa aattgaggtc atataagccc agaagactat ttcagtctgt
     5101 taaacttcat tgccctaaat gtcatttgct gcaagaagtt ccacatgagg gcgatttgga
     5161 tataattttt caggatggtg caactaaaac cccagatgtc aagctacaaa atacatcatt
     5221 atatgattca aaaatctgga ccactaaaaa tcaaaaagga cgaaaagtag cagttcattt
     5281 tgtgaaaaat aatggtattc tcccgctttc aaatgaatgt ctacttttga tagaaggagg
     5341 tacactcagt gaaatttgca aactctcgaa caagtttaat agtgtaattc ctgtgagatc
     5401 tggccacgaa gacctggaac ttttggacct ttcagcacca tttcttatac aaggaacaat
     5461 acatcactat ggatgtaaac agtgttctag tttgagatcc atacaaaatc taaattccct
     5521 ggttgataaa acatcgtgga ttccttcttc tgtggcagaa gcactgggta ttgtacccct
     5581 ccaatatgtg tttgttatga cctttacact tgatgatgga acaggagtac tagaagccta
     5641 tctcatggat tctgacaaat tcttccagat tccagcatca gaagttctga tggatgatga
     5701 ccttcagaaa agtgtggata tgatcatgga tatgttttgt cctccaggaa taaaaattga
     5761 tgcatatccg tggttggaat gcttcatcaa gtcatacaat gtcacaaatg gaacagataa
     5821 tcaaatttgc tatcagattt ttgacaccac agttgcagaa gatgtaatct aagcttgtcg
     5881 agaagtacta gaggatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     5941 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     6001 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     6061 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     6121 atcatgtctg gatctgatca ctgcttgagc ctagaagatc cggctgctaa caaagcccga
     6181 aaggaagctg agttggctgc tgccaccgct gagcaataac tatcataacc cctagggtat
     6241 acccatctaa ttggaaccag ataagtgaaa tctagttcca aactattttg tcatttttaa
     6301 ttttcgtatt agcttacgac gctacaccca gttcccatct attttgtcac tcttccctaa
     6361 ataatcctta aaaactccat ttccacccct cccagttccc aactattttg tccgcccaca
     6421 accggtggag gaaattctcc ttgaagtttc cctggtgttc aaagtaaagg agtttgcacc
     6481 agacgcacct ctgttcactg gtccggcgta ttaaaacacg atacattgtt attagtacat
     6541 ttattaagcg ctagattctg tgcgttgttg atttacagac aattgttgta cgtattttaa
     6601 taattcatta aatttataat ctttagggtg gtatgttaga gcgaaaatca aatgattttc
     6661 agcgtcttta tatctgaatt taaatattaa atcctcaata gatttgtaaa ataggtttcg
     6721 attagtttca aacaagggtt gtttttccga accgatggct ggactatcta atggattttc
     6781 gctcaacgcc acaaaacttg ccaaatcttg tagcagcaat ctagctttgt cgatattcgt
     6841 ttgtgttttg ttttgtaata aaggttcgac gtcgttcaaa atattatgcg cttttgtatt
     6901 tctttcatca ctgtcgttag tgtacaattg actcgacgta aacacgttaa atagagcttg
     6961 gacatattta acatcgggcg tgttagcttt attaggccga tta
//
