LOCUS       sequence-326669-        6575 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ORF68 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6575)
  AUTHORS   Didychuk AL, Gates SN, Gardner MR, Strong LM, Martin A, Glaunsinger 
            BA
  TITLE     A pentameric protein ring with novel architecture is required for 
            herpesviral packaging.
  JOURNAL   Elife. 2021 Feb 8;10. pii: 62261. doi: 10.7554/eLife.62261.
  PUBMED    33554858
REFERENCE   2  (bases 1 to 6575)
  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: "2021-02-8"; volume: "10. pii: 62261. doi"; pages: " 
            10.7554/eLife.62261"
COMMENT     Sequence Label: pCDNA4/TO-ORF68 (tagless)
FEATURES             Location/Qualifiers
     source          1..6575
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(43..62)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        234..613
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        614..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     768..788
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    819..837
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    840..858
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     868..892
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1305..1314
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2473..2496
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2533..2556
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2582..2599)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2588..2812
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2858..3286
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2945..2964)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3155..3176
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3295..3315)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3300..3629
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3480..3615
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3542..3561
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3677..3724
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3743..4117
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4247..4368
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4284..4303)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4338..4357
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4417..4433)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4417..4433)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4430..4452)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4441..4457
                     /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(4465..4471,4472..4489,4490..4495))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4510..4531
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4648..4665)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4819..5407)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4899..4918)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5578..6369,6370..6438))
                     /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     6201..6220
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6439..6543)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
       61 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
      121 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
      181 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
      241 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      301 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      361 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      421 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      481 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      541 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      601 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      661 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      721 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      781 taggcgtgta cggtgggagg tctatataag cagagctctc cctatcagtg atagagatct
      841 ccctatcagt gatagagatc gtcgacgagc tcgtttagtg aaccgtcaga tcgcctggag
      901 acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca gcctccggac
      961 tctagcgttt aaacttaagc ttggtaccga gctcggatcc actagtccag tgtggtggaa
     1021 ttctgcagat atgtttgttc cctggcaact cggtacaatt acccgtcacc gagatgagct
     1081 ccaaaaacta ctggcagcct ccctgctccc ggagcacccg gaggagagcc tcggtaaccc
     1141 cataatgaca cagattcacc agtcgctcca accatcttcc ccctgcaggg tctgtcagct
     1201 cctattttct ctggtccgcg attcgtccac ccccatgggt ttcttcgagg actatgcctg
     1261 cctctgcttc ttctgtctat acgccccaca ctgctggacc tcgaccatgg cggcagcggc
     1321 agacctgtgc gagatcatgc atctgcactt tccagaagag gaggcgacat acgggctatt
     1381 cggaccgggt cgccttatgg gtatcgactt gcagctgcac ttctttgttc aaaagtgctt
     1441 taagaccacc gccgccgaaa aaatactggg aatatccaac ctgcaatttt taaaatcaga
     1501 attcatccgg ggcatgctca caggcaccat cacctgcaac ttctgcttca aaacgtcctg
     1561 gcccaggaca gacaaggagg aggccaccgg ccccacccca tgctgccaga ttacagacac
     1621 caccaccgca cccgcgagcg gcataccgga actagcccgg gccacattct gcggcgcaag
     1681 tcgccccaca aagcccagcc tacttcccgc gctaatagat atctggtcca cgagctcaga
     1741 gctccttgac gagccgcgcc ctcgactgat cgcaagcgac atgagtgaac tcaaatccgt
     1801 ggtcgcatcc cacgatccgt tcttctctcc cccgcttcag gcagacacct cccagggtcc
     1861 atgtctaatg cacccaaccc tggggctacg atacaaaaac gggactgcat ccgtctgcct
     1921 cctctgcgag tgccttgcgg cacacccaga ggcacccaag gcgctgcaga cccttcagtg
     1981 cgaggtaatg ggccatatag aaaacaacgt aaagctggta gacagaattg cctttgtgtt
     2041 ggacaaccca ttcgccatgc catatgtctc agatccgcta cttagagagc tgatccgggg
     2101 ctgtacccca caggaaattc acaagcacct gttctgcgac ccgctgtgcg ccctcaatgc
     2161 taaggtggtg tcagaggacg tactattccg cctgcccagg gagcaggagt ataaaaagct
     2221 cagggcatcc gcggccgccg gacagctcct cgatgccaac accctgttcg actgcgaggt
     2281 cgtgcagact ttggtctttc tctttaaggg tctccaaaac gccagggtgg ggaaaaccac
     2341 ctcactagac attattcggg agctaaccgc acaactaaaa agacaccgcc tagacctggc
     2401 ccacccctca cagacgtcac acttgtacgc ttgaatccag cacagtggcg gccgctcgag
     2461 ggaggcggtg gatggagcca cccccagttc gagaagggcg gcggcagcgg cggcggcagc
     2521 ggcggcggca gctggagcca cccccagttc gagaaggggc ccgtttaaac ccgctgatca
     2581 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     2641 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     2701 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     2761 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     2821 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     2881 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2941 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3001 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3061 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3121 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3181 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3241 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     3301 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3361 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3421 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3481 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3541 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     3601 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     3661 gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata
     3721 cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc
     3781 gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt
     3841 cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt
     3901 ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga
     3961 gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc
     4021 catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg
     4081 caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc
     4141 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4201 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     4261 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4321 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     4381 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4441 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4501 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4561 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4621 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4681 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4741 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4801 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4861 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4921 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4981 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5041 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5101 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5161 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5221 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5281 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5341 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5401 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5461 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5521 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5581 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5641 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5701 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5761 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5821 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5881 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5941 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6001 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6061 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6121 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6181 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6241 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6301 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6361 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6421 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6481 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6541 gcgcacattt ccccgaaaag tgccacctga cgtcg
//
