LOCUS       sequence-486275-        3915 bp    DNA     circular SYN 16-SEP-2025
DEFINITION  BsaI Golden Gate assembly plasmid encoding the SynZiFTR ZF10-NS3-p65
            CDS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3915)
  AUTHORS   Peterman EL, Ploessl DS, Love KS, Sanabria V, Daniels RF, Johnstone 
            CP, Godavarti DR, Kabaria SR, Oakes CG, Pai AA, Galloway KE
  TITLE     High-resolution profiling reveals coupled transcriptional and 
            translational regulation of transgenes.
  JOURNAL   Nucleic Acids Research, Volume 53, Issue 11, 24 June 2025, gkaf528
  PUBMED    39651241
REFERENCE   2  (bases 1 to 3915)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Research, Volume 53, Issue 11, 24 June 2025, gkaf528"
COMMENT     Sequence Label: pKG2197_pPV2-ZF10-NS3-p65
FEATURES             Location/Qualifiers
     source          1..3915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      49..637
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     538..557
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1209..1274
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1281..1301
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2151..2177
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2403..2774
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALLSQISS"
     promoter        2828..2932
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2933..3001,3002..3793)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGYRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3151..3170)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
       61 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
      121 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
      181 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
      241 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
      301 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
      361 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
      421 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
      481 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
      541 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
      601 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg ggtctcaaat
      661 gagccaccat gtctagaccc ggagagcgcc cattccagtg tcggatttgc atgcggaact
      721 tttcgagaag acacggcctg gacagacata cccgtactca tacaggtgaa aaaccctttc
      781 agtgtcggat ctgtatgcga aatttctccg accacagcag cctgaagaga catctacgta
      841 cccacaccgg cagccagaag ccatttcagt gtcggatctg tatgcggaac ttctccgtga
      901 gacacaacct gaccagacat ctacgtacgc acaccggaga gaagccattc caatgccgaa
      961 tatgcatgcg caacttcagt gaccacagca acctgagcag acacctaaaa acccacaccg
     1021 gttcccagaa gccatttcag tgtcggatct gtatgcggaa cttctcccag cgcagcagcc
     1081 tggtgagaca tctacgtacg cacaccggag agaagccatt ccaatgccga atatgcatgc
     1141 gcaacttcag tgagagcggc cacctgaaga gacacctgcg tacgcacctg aggggatcca
     1201 cctgcaggga ctacaaagac catgacggtg attataaaga tcatgacatc gattacaagg
     1261 atgacgatga caagatggcc cccaagaaaa agaggaaggt gggcattcac ggggtgccgg
     1321 gtggactcga gggaggcggt ggaagcggcg gtaccgagga cgtggtgtgc tgccactcaa
     1381 tctacggcaa gaagaagggt gatatcgaca cctaccgata cataggctct tccgggacag
     1441 gctgcgtggt catagtgggc aggatcgtct tgtccggatc cggcactagt gcgcccatca
     1501 cggcgtacgc ccagcagacg agaggcctcc tagggtgtat aatcaccagc ctgactggcc
     1561 gggacaaaaa ccaagtggag ggtgaggtcc agatcgtgtc aactgctacc caaaccttcc
     1621 tggcaacgtg catcaatggg gtatgctggg cagtctacca cggggccgga acgaggacca
     1681 tcgcatcacc caagggtcct gtcatccaga tgtataccaa tgtggaccaa gaccttgtgg
     1741 gctggcccgc tcctcaaggt tcccgctcat tgacaccctg tacctgcggc tcctcggacc
     1801 tttacctggt cacgaggcac gccgatgtca ttcccgtgcg ccggcgaggt gatagcaggg
     1861 gtagcctgct ttcgccccgg cccatttcct acttgaaagg ctcctcgggg ggtccgctgt
     1921 tgtgccccgc gggacacgcc gtgggcctat tcagggccgc ggtgtgcacc cgtggagtgg
     1981 ctaaagcggt ggactttatc cctgtggaga acctagagac aaccatgaga tccccggtgt
     2041 tcacggacaa ctcctctcca ccagcagtca ccctgacgca cccaatcacc aaaatcgata
     2101 gggaggttct ctaccaggag ttcgatgaga tggaagagtg ctctcagcac tatccctacg
     2161 atgtgcccga ttacgctgga ggcggtggaa gcggcggtac cgatgagttt cccaccatgg
     2221 tgtttccttc tgggcagatc agccaggcct cggccttggc cccggcccct ccccaagtcc
     2281 tgccccaggc tccagcccct gcccctgctc cagccatggt atcagctctg gcccaggccc
     2341 cagcccctgt cccagtccta gccccaggcc ctcctcaggc tgtggcccca cctgccccca
     2401 agcccaccca ggctggggaa ggaacgctgt cagaggccct gctgcagctg cagtttgatg
     2461 atgaagacct gggggccttg cttggcaaca gcacagaccc agctgtgttc acagacctgg
     2521 catccgtcga caactccgag tttcagcagc tgctgaacca gggcatacct gtggcccccc
     2581 acacaactga gcccatgctg atggagtacc ctgaggctat aactcgccta gtgacagggg
     2641 cccagaggcc ccccgaccca gctcctgctc cactgggggc cccggggctc cccaatggcc
     2701 tcctttcagg agatgaagac ttctcctcca ttgcggacat ggacttctca gccctgctga
     2761 gtcagatcag ctcccaattg taacaacaga gacctagacg tcaggtggca cttttcgggg
     2821 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     2881 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     2941 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3001 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3061 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3121 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
     3181 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3241 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     3301 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     3361 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     3421 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     3481 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     3541 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     3601 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt atcgcggtat
     3661 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     3721 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     3781 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     3841 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     3901 cccttaacgt gagtt
//
