LOCUS       sequence-257291-        5539 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses the KHSV LANA promoter driving puromycin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5539)
  AUTHORS   Hartenian EN, Glaunsinger BA
  TITLE     Xrn1 activity broadly represses RNA polymerase II occupancy at 
            mammalian but not viral promoters during herpesvirus infection
  JOURNAL   bioRxiv 585984
REFERENCE   2  (bases 1 to 5539)
  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: "bioRxiv 
            585984"
COMMENT     Sequence Label: pGL3-kLANA-puro
FEATURES             Location/Qualifiers
     source          1..5539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             636..1235
                     /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(636..655)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1132..1152
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    1275..1396
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1286..1305)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1340..1359
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    1621..1642
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1657..1662,1663..1680,1681..1687)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1695..1711
                     /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     1700..1722
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1719..1735
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1719..1735
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1753..1771
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1753..1770
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     RBS             2445..2453
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(2684..2703)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2685..2703)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2710..2727)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2710..2726)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2719..2741)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2871..2888)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3042..3630)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3122..3141)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3801..4592,4593..4661))
                     /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     4424..4443
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4662..4766)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4793..5248
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4880..4899)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5090..5111
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5379..5427
                     /note="synthetic polyadenylation signal"
     misc_feature    5441..5532
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5481..5500
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgaggct agccagatga acgccaccca
       61 aggtaatgaa atgaagaggc tagacatgga cccccgagag gcactcggcg tcgtccacgg
      121 gggagcacag acgacaggtc ggaccactga accggtgcca gcaaccggga gcacagtggg
      181 gggtaaattc tgcaaagcag acacgccttc ttcagttttt atggcagtcc tgcctggggc
      241 accaatcaga aagtagcttg atataactcc gccctccact acgcggcgcc cgggacaatc
      301 aaaaaatgtg tgtatcattt ggaggcagct gcgccacgaa gcagtcacgt ccccaagagc
      361 agcagcttgg tccggctgac ttataaacag tgagtagcgc ccgcccactt gtttcgggcg
      421 gcccgtaagg cacccttatc tttgaaatgc tataagatgt gtgggaaata gtaataccag
      481 gtgtacatga tttgtgttaa ggtgtgtggt ggttttcgaa aaacatgggt tattggccgt
      541 ttctgtttct cacgcccgga ttatatatct ggacttgcct aggtagcatc catggaagct
      601 tggcattccg gtactgttgg ttaactataa cggtcatgac cgagtacaag cccacggtgc
      661 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
      721 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
      781 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
      841 acggcgccgc cgtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
      901 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
      961 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     1021 gagtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     1081 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     1141 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     1201 gcacctggtg catgacccgc aagcccggtg cctgatctag agtcggggcg gccggccgct
     1261 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     1321 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     1381 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     1441 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga
     1501 tccgtcgacc gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg
     1561 gcatgactac tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     1621 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     1681 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     1741 ttacgccaag ctatttaggt gacactatag aatactcaag ctatgcatcc aacgcgttgg
     1801 gagctctccg gatccaagct ggaaccaaca gcgcccctaa aatcggtttg cggttagacc
     1861 aggcaaaaac acatcaaaaa gtagcacttt aagcggcctc tgccgcatcg cctcacagaa
     1921 aaagcgcatt acgtcattta aaaaaacaca tcaaaaagtg ataacttgat gtgtgggtgt
     1981 ggtttacagg cacacaaagg ccacaccccc ggctcccgtc aagttggtga ttggtccgtc
     2041 aaaaaatcaa tcaaggaatc cggactactg attggttacg cccaaaaatc cgacccagcc
     2101 attggataaa acgtgctagc caatcaggtg ggaggcggat ccctataata tatagtatta
     2161 tttatttatt aatgtagggt accttttggt gggaccaata agtatgccta tcctgtaaca
     2221 cgtaaaaggt agggtgtgga tagttggcaa taggccaggt gggccgaacc attaaggttt
     2281 aaccaattag gatagaccct ctcataaacc actcccattt tacacatcct ccctctgttt
     2341 agggggatgc atatggttgg gttcaccctg ggggtcccca tattgaatat accttggttt
     2401 cccttttgac tgttgccatg acaactatgg tatgtcaacc cctgacctcc ttacctgttg
     2461 ataccttttg cggtttttat gaatcacctg ttggtagttt aaaagtacac tttttgatgt
     2521 gtttttctta accgaggtta tatatagacc ctgatttctg tgggggctca ttctgcgccg
     2581 cgcctcgcta ctggacggac ctctgctgtg ctgctcctcg ctttggctcc ccaggccggg
     2641 ggacccgcac cccgggccgg gcagcccgct ccggcccaat tcgccctata gtgagtcgta
     2701 ttacaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     2761 acttaatcgc cttgcagcac atcccccttt cgccagtcgt cgccgcactt atgactgtct
     2821 tctttatcat gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact
     2881 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2941 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3001 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3061 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3121 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3181 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3241 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3301 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3361 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3421 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3481 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3541 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3601 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3661 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3721 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3781 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3841 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3901 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3961 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4021 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4081 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4141 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4201 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4261 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4321 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4381 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4441 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4501 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4561 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4621 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     4681 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4741 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc
     4801 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4861 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4921 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4981 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     5041 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     5101 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     5161 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     5221 tttaacaaaa tattaacgct tacaatttgc cattcgccat tcaggctgcg caactgttgg
     5281 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc ccaagctacc atgataagta
     5341 agtaatatta aggtacggga ggtacttgga gcggccgcaa taaaatatct ttattttcat
     5401 tacatctgtg tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa
     5461 acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca
     5521 gaacatttct ctatcgata
//
