LOCUS       sequence-469013-        7942 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  240 basepair upstream sequence of KSHV ORF68 driving HaloTag 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7942)
  AUTHORS   Morgens DW, Gulyas L, Mao X, Rivera-Madera A, Souza AS, Glaunsinger 
            BA
  TITLE     Enhancers and genome conformation provide complex transcriptional 
            control of a herpesviral gene.
  JOURNAL   Mol Syst Biol. 2025 Jan;21(1):30-58. doi: 
            10.1038/s44320-024-00075-0. Epub 2024 Nov 19.
  PUBMED    39562742
REFERENCE   2  (bases 1 to 7942)
  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: 
            "10.1038/s44320-024-00075-0"; journalName: "Mol Syst Biol"; date: 
            "2025-01"; volume: "21"; issue: "1"; pages: "30-58"
COMMENT     Sequence Label: pGlab592
FEATURES             Location/Qualifiers
     source          1..7942
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             80..313
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    385..506
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(422..441)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     476..495
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      546..681
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(701..720)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(702..720)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(730..747)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(730..746)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(739..761)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      888..1343
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(975..994)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1185..1206
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1369..1473
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1474..1542,1543..2334)
                     /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(1692..1711)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2505..3093
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2994..3013
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3247..3264
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3381..3402
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3417..3422,3423..3440,3441..3447)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3455..3471
                     /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     3460..3482
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3479..3495
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3479..3495
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3514..3534
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3516..3534
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3562..3788
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             3789..3969
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4016..4141
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4634..4867
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5052..5096
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     CDS             5611..6498
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="AEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
     CDS             6532..6552
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    6597..6714
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6763..7273
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7192..7210
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             7295..7894
                     /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     complement(7295..7314)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7791..7811
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 catgcatcgg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
       61 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt
      121 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
      181 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
      241 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
      301 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag
      361 aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa
      421 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
      481 ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc
      541 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
      601 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
      661 gaggaggctt ttttggaggc ctagggacgt acccaattcg ccctatagtg agtcgtatta
      721 cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
      781 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
      841 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
      901 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
      961 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     1021 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     1081 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     1141 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     1201 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     1261 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     1321 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
     1381 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     1441 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     1501 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     1561 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     1621 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     1681 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     1741 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     1801 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     1861 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     1921 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     1981 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     2041 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     2101 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     2161 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     2221 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     2281 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     2341 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     2401 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     2461 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     2521 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     2581 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     2641 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     2701 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     2761 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     2821 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     2881 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     2941 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     3001 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     3061 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     3121 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     3181 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     3241 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     3301 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     3361 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcaac ccaggcttta
     3421 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     3481 ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa
     3541 aagctggagc tgcaagctta atgtagtctt atgcaatact cttgtagtct tgcaacatgg
     3601 taacgatgag ttagcaacat gccttacaag gagagaaaaa gcaccgtgca tgccgattgg
     3661 tggaagtaag gtggtacgat cgtgccttat taggaaggca acagacgggt ctgacatgga
     3721 ttggacgaac cactgaattg ccgcattgca gagatattgt atttaagtgc ctagctcgat
     3781 acataaacgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     3841 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     3901 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     3961 tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc
     4021 tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt
     4081 gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt
     4141 cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg
     4201 aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc
     4261 agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca
     4321 accatccctt cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct
     4381 ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag acaagataga
     4441 ggaagagcaa aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg
     4501 gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa
     4561 ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa
     4621 gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg
     4681 gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc
     4741 agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct
     4801 ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac
     4861 agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga
     4921 atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt
     4981 gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa
     5041 accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga
     5101 attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag
     5161 gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg
     5221 gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg
     5281 aaggaataga agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg
     5341 gatctcgacg gtatcgatca cgagactagc tcacgaggca gaagctggtc agacgcgtac
     5401 tccatgttgg gttccacggg gaggggaaca cacgccccaa gacacgacgg cgcacatagg
     5461 gagcggagca aacaattgat tcaaatattt gactccgcag cgagccggtt tgcagagtgg
     5521 tcacctgccc tgctccacac ccacccccgc gtctcttcca actctcaact cacgatccag
     5581 ggaaaccacc gtccagtggc catgtttgtt gcagaaatcg gtactggctt tccattcgac
     5641 ccccattatg tggaagtcct gggcgagcgc atgcactacg tcgatgttgg tccgcgcgat
     5701 ggcacccctg tgctgttcct gcacggtaac ccgacctcct cctacgtgtg gcgcaacatc
     5761 atcccgcatg ttgcaccgac ccatcgctgc attgctccag acctgatcgg tatgggcaaa
     5821 tccgacaaac cagacctggg ttatttcttc gacgaccacg tccgcttcat ggatgccttc
     5881 atcgaagccc tgggtctgga agaggtcgtc ctggtcattc acgactgggg ctccgctctg
     5941 ggtttccact gggccaagcg caatccagag cgcgtcaaag gtattgcatt tatggagttc
     6001 atccgcccta tcccgacctg ggacgaatgg ccagaatttg cccgcgagac cttccaggcc
     6061 ttccgcacca ccgacgtcgg ccgcaagctg atcatcgatc agaacgtttt tatcgagggt
     6121 acgctgccga tgggtgtcgt ccgcccgctg actgaagtcg agatggacca ttaccgcgag
     6181 ccgttcctga atcctgttga ccgcgagcca ctgtggcgct tcccaaacga gctgccaatc
     6241 gccggtgagc cagcgaacat cgtcgcgctg gtcgaagaat acatggactg gctgcaccag
     6301 tcccctgtcc cgaagctgct gttctggggc accccaggcg ttctgatccc accggccgaa
     6361 gccgctcgcc tggccaaaag cctgcctaac tgcaaggctg tggacatcgg cccgggtctg
     6421 aatctgctgc aagaagacaa cccggacctg atcggcagcg agatcgcgcg ctggctgtcg
     6481 acgctcgaga tttccggatc cgctggctcc gctgctggtt ctggcgaatt cccaaagaag
     6541 aagcggaagg tctgagaatt ctcgacctcg agacaaatgg cagtattcat ccacaatttt
     6601 aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca
     6661 acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt
     6721 tattacaggg acagcagaga tccactttgg ccgcggctcg agggggttgg ggttgcgcct
     6781 tttccaaggc agccctgggt ttgcgcaggg acgcggctgc tctgggcgtg gttccgggaa
     6841 acgcagcggc gccgaccctg ggtctcgcac attcttcacg tccgttcgca gcgtcacccg
     6901 gatcttcgcc gctacccttg tgggcccccc ggcgacgctt cctgctccgc ccctaagtcg
     6961 ggaaggttcc ttgcggttcg cggcgtgccg gacgtgacaa acggaagccg cacgtctcac
     7021 tagtaccctc gcagacggac agcgccaggg agcaatggca gcgcgccgac cgcgatgggc
     7081 tgtggccaat agcggctgct cagcagggcg cgccgagagc agcggccggg aaggggcggt
     7141 gcgggaggcg gggtgtgggg cggtagtgtg ggccctgttc ctgcccgcgc ggtgttccgc
     7201 attctgcaag cctccggagc gcacgtcggc agtcggctcc ctcgttgacc gaatcaccga
     7261 cctctctccc cagggggatc caccggagct taccatgacc gagtacaagc ccacggtgcg
     7321 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     7381 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     7441 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     7501 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     7561 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     7621 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     7681 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     7741 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     7801 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     7861 cacctggtgc atgacccgca agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc
     7921 gaccgaaagg agcgcacgac cc
//
