LOCUS       sequence-447352-        5473 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for an inducible V5-tagged luciferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5473)
  TITLE     Janes lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5473)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pEN_TT luciferase-V5
FEATURES             Location/Qualifiers
     source          1..5473
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(446..979)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1168..1173,1174..1190,1191..1196))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      1853..2441
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2342..2361
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      2606..2692
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2784..2811
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     2819..2842
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    2861..2960
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2992..3306
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    3000..3018
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3036..3054
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3071..3089
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3107..3125
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3143..3161
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3178..3196
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3214..3232
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3279..3303
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     complement(3379..3397)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     CDS             4957..4998
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    complement(5037..5136)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5248..5267)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(5326..5345)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
       61 atggttactc accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc
      121 tgattcaggt gaaaatattg ttgatgcgct ggcagtgtcc ctgcgccggt tgcattcgat
      181 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
      241 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
      301 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
      361 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taggccggga
      421 agccgatctc ggcttgaacg aattgttagg tggcggtact tgggtcgata tcaaagtgca
      481 tcacttcttc ccgtatgccc aactttgtat agagagccac tgcgggatcg tcaccgtaat
      541 ctgcttgcac gtagatcaca taagcaccaa gcgcgttggc ctcatgcttg aggagattga
      601 tgagcgcggt ggcaatgccc tgcctccggt gctcgccgga gactgcgaga tcatagatat
      661 agatctcact acgcggctgc tcaaacttgg gcagaacgta agccgcgaga gcgccaacaa
      721 ccgcttcttg gtcgaaggca gcaagcgcga tgaatgtctt actacggagc aagttcccga
      781 ggtaatcgga gtccggctga tgttgggagt aggtggctac gtctccgaac tcacgaccga
      841 aaagatcaag agcagcccgc atggatttga cttggtcagg gccgagccta catgtgcgaa
      901 tgatgcccat acttgagcca cctaactttg ttttagggcg actgccctgc tgcgtaacat
      961 cgttgctgct gcgtaacatc gttgctgctc cataacatca aacatcgacc cacggcgtaa
     1021 cgcgcttgct gcttggatgc ccgaggcata gactgtacaa aaaaacagtc ataacaagcc
     1081 atgaaaaccg ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct ggaccagttg
     1141 cgtgagcgca tacgctactt gcattacagt ttacgaaccg aacaggctta tgtcaactgg
     1201 gttcgtgcct tcatccgttt ccacggtgtg cgtcacccgg caaccttggg cagcagcgaa
     1261 gtcgaggcat ttctgtcctg gctggcgaac gagcgcaagg tttcggtctc cacgcatcgt
     1321 caggcattgg cggccttgct gttcttctac ggcaaggtgc tgtgcacgga tctgccctgg
     1381 cttcaggaga tcggaagacc tcggccgtcg cggcgcttgc cggtggtgct gaccccggat
     1441 gaagtggttc gcatcctcgg ttttctggaa ggcgagcatc gtttgttcgc ccaggactct
     1501 agctatagtt ctagtggttg gctactaata ggttgtattg atgttggacg agtcggaatc
     1561 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     1621 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     1681 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg
     1741 cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg
     1801 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     1861 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     1921 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     1981 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     2041 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     2101 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     2161 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     2221 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     2281 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     2341 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     2401 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     2461 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     2521 ctgattctgt ggataaccgt attaccgcta gcatggatct cggggacgtc taactactaa
     2581 gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcgg aagactgggc
     2641 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg
     2701 agcggatttg aacgttgtga agcaacggcc cggagggtgg cgggcaggac gcccgccata
     2761 aactgccagg catcaaacta agcagaaggc catcctgacg gatggccttt ttgcgtttct
     2821 acaaactctt cctgttagtt agttacttaa gctcgggccc caaataatga ttttattttg
     2881 actgatagtg acctgttcgt tgcaacaaat tgataagcaa tgctttttta taatgccaac
     2941 tttgtacaaa aaagcaggct ttaaaggaac caattcagtc gactggatcc cgagtttact
     3001 ccctatcagt gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgatg
     3061 tcgagtttac tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat
     3121 agagaacgta tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttatcc
     3181 ctatcagtga tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgtatgt
     3241 cgaggtaggc gtgtacggtg ggaggcctat ataagcagag ctcgtttagt gaaccgtcag
     3301 atcgccacta gtatggaaga cgccaaaaac ataaagaaag gcccggcgcc attctatccg
     3361 ctggaagatg gaaccgctgg agagcaactg cataaggcta tgaagagata cgccctggtt
     3421 cctggaacaa ttgcttttac agatgcacat atcgaggtgg acatcactta cgctgagtac
     3481 ttcgaaatgt ccgttcggtt ggcagaagct atgaaacgat atgggctgaa tacaaatcac
     3541 agaatcgtcg tatgcagtga aaactctctt caattcttta tgccggtgtt gggcgcgtta
     3601 tttatcggag ttgcagttgc gcccgcgaac gacatttata atgaacgtga attgctcaac
     3661 agtatgggca tttcgcagcc taccgtggtg ttcgtttcca aaaaggggtt gcaaaaaatt
     3721 ttgaacgtgc aaaaaaagct cccaatcatc caaaaaatta ttatcatgga ttctaaaacg
     3781 gattaccagg gatttcagtc gatgtacacg ttcgtcacat ctcatctacc tcccggtttt
     3841 aatgaatacg attttgtgcc agagtccttc gatagggaca agacaattgc actgatcatg
     3901 aactcctctg gatctactgg tctgcctaaa ggtgtcgctc tgcctcatag aactgcctgc
     3961 gtgagattct cgcatgccag agatcctatt tttggcaatc aaatcattcc ggatactgcg
     4021 attttaagtg ttgttccatt ccatcacggt tttggaatgt ttactacact cggatatttg
     4081 atatgtggat ttcgagtcgt cttaatgtat agatttgaag aagagctgtt tctgaggagc
     4141 cttcaggatt acaagattca aagtgcgctg ctggtgccaa ccctattctc cttcttcgcc
     4201 aaaagcactc tgattgacaa atacgattta tctaatttac acgaaattgc ttctggtggc
     4261 gctcccctct ctaaggaagt cggggaagcg gttgccaaga ggttccatct gccaggtatc
     4321 aggcaaggat atgggctcac tgagactaca tcagctattc tgattacacc cgagggggat
     4381 gataaaccgg gcgcggtcgg taaagttgtt ccattttttg aagcgaaggt tgtggatctg
     4441 gataccggga aaacgctggg cgttaatcaa agaggcgaac tgtgtgtgag aggtcctatg
     4501 attatgtccg gttatgtaaa caatccggaa gcgaccaacg ccttgattga caaggatgga
     4561 tggctacatt ctggagacat agcttactgg gacgaagacg aacacttctt catcgttgac
     4621 cgcctgaagt ctctgattaa gtacaaaggc tatcaggtgg ctcccgctga attggaatcc
     4681 atcttgctcc aacaccccaa catcttcgac gcaggtgtcg caggtcttcc cgacgatgac
     4741 gccggtgaac ttcccgccgc cgttgttgtt ttggagcacg gaaagacgat gacggaaaaa
     4801 gagatcgtgg attacgtcgc cagtcaagta acaaccgcga aaaagttgcg cggaggagtt
     4861 gtgtttgtgg acgaagtacc gaaaggtctt accggaaaac tcgacgcaag aaaaatcaga
     4921 gagatcctca taaaggccaa gaagggcgga aagatcggta agcctatccc taaccctctc
     4981 ctcggtctcg attctacgtg agaattggaa gactaatgcg ttttcgagat atctagaccc
     5041 agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga
     5101 acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgcag ctctggcccg
     5161 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa
     5221 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa
     5281 cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     5341 gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc
     5401 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     5461 tcagactaaa ctg
//
