LOCUS       sequence-12076-e        4347 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector with TRE promoter followed by multiple cloning site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4347)
  AUTHORS   Shin KJ, Wall EA, Zavzavadjian JR, Santat LA, Liu J, Hwang JI, 
            Rebres R, Roach T, Seaman W, Simon MI, Fraser ID
  TITLE     A single lentiviral vector platform for microRNA-based conditional 
            RNA interference and coordinated transgene expression.
  JOURNAL   Proc Natl Acad Sci U S A. 2006 Sep 12. 103(37):13759-64.
  PUBMED    16945906
REFERENCE   2  (bases 1 to 4347)
  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: "Proc Natl 
            Acad Sci U S A. 2006 Sep 12. 103(37):13759-64."
COMMENT     Sequence Label: pEN_Tmcs
FEATURES             Location/Qualifiers
     source          1..4347
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     25..48
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    67..166
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    join(205..223,224..246,247..265,266..288,289..307,308..330,
                     331..349,350..372,373..391,392..414,415..433,434..456,
                     457..475)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        508..546
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     543..567
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             783..1088
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRIVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     967..986
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1117..1216)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1328..1347)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1406..1425)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(1999..2532)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2721..2726,2727..2743,2744..2749))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      3406..3994
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3895..3914
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      4159..4245
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(4337..4347,1..17)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ctgacggatg gcctttttgc gtttctacaa actcttcctg ttagttagtt acttaagctc
       61 gggccccaaa taatgatttt attttgactg atagtgacct gttcgttgca acaaattgat
      121 aagcaatgct tttttataat gccaactttg tacaaaaaag caggctttaa aggaaccaat
      181 tcagtcgact ggatccttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt
      241 taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg
      301 atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt
      361 cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta
      421 tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt
      481 gaaagtcgag ctcggtaccc gggtcgaggt aggcgtgtac ggtgggaggc ctatataagc
      541 agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc
      601 catagaagac accgggaccg atccagcctc cgcggccccg aactagtcga ctggatccgg
      661 taccgaattc gcttactaaa agccagataa cagtatgcgt atttgcgcgc tgatttttgc
      721 ggtataagaa tatatactga tatgtatacc cgaagtatgt caaaaagagg tgtgcttcta
      781 gaatgcagtt taaggtttac acctataaaa gagagagccg ttatcgtctg tttgtggatg
      841 tacagagtga tattattgac acgcccgggc gacggatagt gatccccctg gccagtgcac
      901 gtctgctgtc agataaagtc tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa
      961 gctggcgcat gatgaccacc gatatggcca gtgtgccggt ctccgttatc ggggaagaag
     1021 tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg atgttctggg
     1081 gaatatagaa ttcgcggccg cactcgagat atctagaccc agctttcttg tacaaagttg
     1141 gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa
     1201 ataaaatcat tatttgccat ccagctgcag ctctggcccg tgtctcaaaa tctctgatgt
     1261 tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac
     1321 agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc gcgattaaat
     1381 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
     1441 ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
     1501 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
     1561 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
     1621 ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata tcctgattca
     1681 ggtgaaaata ttgttgatgc gctggcagtg tccctgcgcc ggttgcattc gattcctgtt
     1741 tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg
     1801 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
     1861 caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat
     1921 ggtgatttct cacttgataa ccttattttt gacgagggga aattaggccg ggaagccgat
     1981 ctcggcttga acgaattgtt aggtggcggt acttgggtcg atatcaaagt gcatcacttc
     2041 ttcccgtatg cccaactttg tatagagagc cactgcggga tcgtcaccgt aatctgcttg
     2101 cacgtagatc acataagcac caagcgcgtt ggcctcatgc ttgaggagat tgatgagcgc
     2161 ggtggcaatg ccctgcctcc ggtgctcgcc ggagactgcg agatcataga tatagatctc
     2221 actacgcggc tgctcaaacc tgggcagaac gtaagccgcg agagcgccaa caaccgcttc
     2281 ttggtcgaag gcagcaagcg cgatgaatgt cttactacgg agcaagttcc cgaggtaatc
     2341 ggagtccggc tgatgttggg agtaggtggc tacgtctccg aactcacgac cgaaaagatc
     2401 aagagcagcc cgcatggatt tgacttggtc agggccgagc ctacatgtgc gaatgatgcc
     2461 catacttgag ccacctaact ttgttttagg gcgactgccc tgctgcgtaa catcgttgct
     2521 gctgcgtaac atcgttgctg ctccataaca tcaaacatcg acccacggcg taacgcgctt
     2581 gctgcttgga tgcccgaggc atagactgta caaaaaaaca gtcataacaa gccatgaaaa
     2641 ccgccactgc gccgttacca ccgctgcgtt cggtcaaggt tctggaccag ttgcgtgagc
     2701 gcatacgcta cttgcattac agtttacgaa ccgaacaggc ttatgtcaac tgggttcgtg
     2761 ccttcatccg tttccacggt gtgcgtcacc cggcaacctt gggcagcagc gaagtcgagg
     2821 catttctgtc ctggctggcg aacgagcgca aggtttcggt ctccacgcat cgtcaggcat
     2881 tggcggcctt gctgttcttc tacggcaagg tgctgtgcac ggatctgccc tggcttcagg
     2941 agatcggaag acctcggccg tcgcggcgct tgccggtggt gctgaccccg gatgaagtgg
     3001 ttcgcatcct cggttttctg gaaggcgagc atcgtttgtt cgcccaggac tctagctata
     3061 gttctagtgg ttggctacta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
     3121 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
     3181 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
     3241 tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca
     3301 ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttttc
     3361 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3421 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3481 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3541 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3601 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3661 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3721 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3781 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3841 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3901 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     3961 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4021 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     4081 tgtggataac cgtattaccg ctagcatgga tctcggggac gtctaactac taagcgagag
     4141 tagggaactg ccaggcatca aataaaacga aaggctcagt cggaagactg ggcctttcgt
     4201 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat
     4261 ttgaacgttg tgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc
     4321 aggcatcaaa ctaagcagaa ggccatc
//
