LOCUS       sequence-183535-        4263 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for inducible expression of human ID2 with 
            N-terminal 3xFLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4263)
  AUTHORS   Bajikar SS, Wang CC, Borten MA, Pereira EJ, Atkins KA, Janes KA
  TITLE     Tumor-Suppressor Inactivation of GDF11 Occurs by Precursor 
            Sequestration in Triple-Negative Breast Cancer.
  JOURNAL   Dev Cell. 2017 Nov 20;43(4):418-435.e13. doi: 
            10.1016/j.devcel.2017.10.027.
  PUBMED    29161592
REFERENCE   2  (bases 1 to 4263)
  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: 
            "10.1016/j.devcel.2017.10"; journalName: "Dev Cell"; date: 
            "2017-11-20- 20"; volume: "43"; issue: "4"; pages: "418-435.e13"
COMMENT     Sequence Label: pEN_TT 3xFLAG ID2
FEATURES             Location/Qualifiers
     source          1..4263
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      102..129
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     137..160
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    179..278
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        310..624
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    318..336
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    354..372
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    389..407
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    425..443
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    461..479
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    496..514
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    532..550
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     597..621
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             633..698
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(1145..1244)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1356..1375)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1434..1453)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(2027..2560)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2749..2754,2755..2771,2772..2777))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      3434..4022
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3923..3942
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      join(4187..4263,1..10)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 taggacaaat ccgccgggag cggatttgaa cgttgtgaag caacggcccg gagggtggcg
       61 ggcaggacgc ccgccataaa ctgccaggca tcaaactaag cagaaggcca tcctgacgga
      121 tggccttttt gcgtttctac aaactcttcc tgttagttag ttacttaagc tcgggcccca
      181 aataatgatt ttattttgac tgatagtgac ctgttcgttg caacaaattg ataagcaatg
      241 cttttttata atgccaactt tgtacaaaaa agcaggcttt aaaggaacca attcagtcga
      301 ctggatcccg agtttactcc ctatcagtga tagagaacgt atgtcgagtt tactccctat
      361 cagtgataga gaacgatgtc gagtttactc cctatcagtg atagagaacg tatgtcgagt
      421 ttactcccta tcagtgatag agaacgtatg tcgagtttac tccctatcag tgatagagaa
      481 cgtatgtcga gtttatccct atcagtgata gagaacgtat gtcgagttta ctccctatca
      541 gtgatagaga acgtatgtcg aggtaggcgt gtacggtggg aggcctatat aagcagagct
      601 cgtttagtga accgtcagat cgccactaga tggactacaa agaccatgac ggtgattata
      661 aagatcatga catcgattac aaggatgacg atgacaagac tagtatgaaa gccttcagtc
      721 ccgtgaggtc cgttaggaaa aacagcctgt cggaccacag cctgggcatc tcccggagca
      781 aaacccctgt ggacgacccg atgagcctgc tatacaacat gaacgactgc tactccaagc
      841 tcaaggagct ggtgcccagc atcccccaga acaagaaggt gagcaagatg gaaatcctgc
      901 agcacgtcat cgactacatc ttggacctgc agatcgccct ggactcgcat cccactattg
      961 tcagcctgca tcaccagaga cccgggcaga accaggcgtc caggacgccg ctgaccaccc
     1021 tcaacacgga tatcagcatc ctgtccttgc aggcttctga attcccttct gagttaatgt
     1081 caaatgacag caaagcactg tgtggctgac aattggaaga ctaatgcgtt ttcgagatat
     1141 ctagacccag ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt
     1201 tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgcagct
     1261 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     1321 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     1381 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     1441 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     1501 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     1561 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     1621 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     1681 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtc
     1741 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     1801 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     1861 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     1921 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     1981 cgaggggaaa ttaggccggg aagccgatct cggcttgaac gaattgttag gtggcggtac
     2041 ttgggtcgat atcaaagtgc atcacttctt cccgtatgcc caactttgta tagagagcca
     2101 ctgcgggatc gtcaccgtaa tctgcttgca cgtagatcac ataagcacca agcgcgttgg
     2161 cctcatgctt gaggagattg atgagcgcgg tggcaatgcc ctgcctccgg tgctcgccgg
     2221 agactgcgag atcatagata tagatctcac tacgcggctg ctcaaacttg ggcagaacgt
     2281 aagccgcgag agcgccaaca accgcttctt ggtcgaaggc agcaagcgcg atgaatgtct
     2341 tactacggag caagttcccg aggtaatcgg agtccggctg atgttgggag taggtggcta
     2401 cgtctccgaa ctcacgaccg aaaagatcaa gagcagcccg catggatttg acttggtcag
     2461 ggccgagcct acatgtgcga atgatgccca tacttgagcc acctaacttt gttttagggc
     2521 gactgccctg ctgcgtaaca tcgttgctgc tgcgtaacat cgttgctgct ccataacatc
     2581 aaacatcgac ccacggcgta acgcgcttgc tgcttggatg cccgaggcat agactgtaca
     2641 aaaaaacagt cataacaagc catgaaaacc gccactgcgc cgttaccacc gctgcgttcg
     2701 gtcaaggttc tggaccagtt gcgtgagcgc atacgctact tgcattacag tttacgaacc
     2761 gaacaggctt atgtcaactg ggttcgtgcc ttcatccgtt tccacggtgt gcgtcacccg
     2821 gcaaccttgg gcagcagcga agtcgaggca tttctgtcct ggctggcgaa cgagcgcaag
     2881 gtttcggtct ccacgcatcg tcaggcattg gcggccttgc tgttcttcta cggcaaggtg
     2941 ctgtgcacgg atctgccctg gcttcaggag atcggaagac ctcggccgtc gcggcgcttg
     3001 ccggtggtgc tgaccccgga tgaagtggtt cgcatcctcg gttttctgga aggcgagcat
     3061 cgtttgttcg cccaggactc tagctatagt tctagtggtt ggctactaat aggttgtatt
     3121 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     3181 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     3241 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     3301 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca
     3361 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     3421 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3481 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3541 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     3601 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3661 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3721 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3781 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3841 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3901 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3961 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     4021 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     4081 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgct agcatggatc
     4141 tcggggacgt ctaactacta agcgagagta gggaactgcc aggcatcaaa taaaacgaaa
     4201 ggctcagtcg gaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
     4261 gag
//
