LOCUS       sequence-375782-        5391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5391)
  AUTHORS   Wang L, Paudel BB, McKnight RA, Janes KA
  TITLE     Nucleocytoplasmic transport of active HER2 causes fractional escape 
            from the DCIS-like state.
  JOURNAL   Nat Commun. 2023 Apr 13;14(1):2110. doi: 10.1038/s41467-023-37914-x.
  PUBMED    37055441
REFERENCE   2  (bases 1 to 5391)
  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: "Nat 
            Commun."; date: "2023-04-13"; volume: "14(1):2110. doi"; pages: " 
            10.1038/s41467-023-37914-x"
COMMENT     Sequence Label: pEN_TT BirA* w ccdB
FEATURES             Location/Qualifiers
     source          1..5391
                     /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"
     promoter        198..512
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    206..224
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    242..260
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    277..295
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    313..331
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    349..367
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    384..402
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    420..438
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     485..509
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(519..869,870..872,873..1481)
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     ncRNA           1513..1634
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     CDS             1670..1975
                     /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     1854..1873
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     ncRNA           1996..2122
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     protein_bind    complement(2161..2260)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2372..2391)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(2450..2469)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(3043..3576)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3765..3770,3771..3787,3788..3793))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      4450..5038
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4939..4958
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      5203..5289
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(5381..5391,1..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 ggatcccgag tttactccct atcagtgata gagaacgtat gtcgagttta
      241 ctccctatca gtgatagaga acgatgtcga gtttactccc tatcagtgat agagaacgta
      301 tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg
      361 atagagaacg tatgtcgagt ttatccctat cagtgataga gaacgtatgt cgagtttact
      421 ccctatcagt gatagagaac gtatgtcgag gtaggcgtgt acggtgggag gcctatataa
      481 gcagagctcg tttagtgaac cgtcagatcg ccactagaat gaaggacaac accgtgcccc
      541 tgaagctgat cgccctgctg gccaacggcg agttccactc tggcgagcag ctgggagaga
      601 ccctgggaat gagcagagcc gccatcaaca agcacatcca gacactgaga gactggggag
      661 tggacgtgtt caccgtgcct ggcaagggct acagcctgcc tgagcctatc cagctgctga
      721 acgccaagca gatcctggga cagctggatg gcggaagcgt ggccgtgctg cctgtgatcg
      781 actccaccaa tcagtacctg ctggacagaa tcggagagct gaagtccggc gacgcctgca
      841 tcgccgagta ccagcaggct ggcagaggag gcagaggacg gaagtggttc agcccattcg
      901 gagccaacct gtacctgtcc atgttctgga gactggagca gggacctgct gctgccatcg
      961 gactgagtct ggtgatcgga atcgtgatgg ccgaggtgct gagaaagctg ggagccgaca
     1021 aggtgagagt gaagtggcct aatgacctgt acctccagga ccgcaagctg gctggcatcc
     1081 tggtggagct gacaggcaag acaggcgatg ccgctcagat cgtgatcgga gccggaatca
     1141 acatggccat gagaagagtg gaggagagcg tggtgaacca gggctggatc accctgcagg
     1201 aggctggcat caacctggac cggaacaccc tggccgccat gctgatcaga gagctgagag
     1261 ccgctctgga gctgttcgag caggagggac tggctcctta cctgagcaga tgggagaagc
     1321 tggacaactt catcaacaga cctgtgaagc tgatcatcgg cgacaaggaa atcttcggca
     1381 tctccagagg aatcgacaag cagggagctc tgctgctgga gcaggacgga atcatcaagc
     1441 cctggatggg cggagaaatc tccctgagaa gcgcagagaa gactagtcga ctagggataa
     1501 cagggtaatt gtttgaatga ggcttcagta ctttacagaa tcgttgcctg cacatcttgg
     1561 aaacacttgc tgggattact tcttcaggtt aacccaacag aaggctcgag aaggtatatt
     1621 gctgttgaca gtgagcgtgc tgcaggtaag cttgaggtgt gcttccagaa tgcagtttaa
     1681 ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac agagtgatat
     1741 tattgacacg cccgggcgac ggatagtgat ccccctggcc agtgcacgtc tgctgtcaga
     1801 taaagtctcc cgtgaacttt acccggtggt gcatatcggg gatgaaagct ggcgcatgat
     1861 gaccaccgat atggccagtg tgccggtctc cgttatcggg gaagaagtgg ctgatctcag
     1921 ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa tatagaattc
     1981 caacctgctg cttgcctact gcctcggaat tcaaggggct actttaggag caattatctt
     2041 gtttactaaa actgaatacc ttgctatctc tttgatacat ttttacaaag ctgaattaaa
     2101 atggtataaa ttaaatcact tttttcaatt ggaagactaa tgcgttttcg agatatctag
     2161 acccagcttt cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca
     2221 acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct gcagctctgg
     2281 cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca
     2341 ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg
     2401 gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     2461 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat
     2521 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     2581 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     2641 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag
     2701 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgtccctg
     2761 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
     2821 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     2881 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc
     2941 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     3001 gggaaattag gccgggaagc cgatctcggc ttgaacgaat tgttaggtgg cggtacttgg
     3061 gtcgatatca aagtgcatca cttcttcccg tatgcccaac tttgtataga gagccactgc
     3121 gggatcgtca ccgtaatctg cttgcacgta gatcacataa gcaccaagcg cgttggcctc
     3181 atgcttgagg agattgatga gcgcggtggc aatgccctgc ctccggtgct cgccggagac
     3241 tgcgagatca tagatataga tctcactacg cggctgctca aacttgggca gaacgtaagc
     3301 cgcgagagcg ccaacaaccg cttcttggtc gaaggcagca agcgcgatga atgtcttact
     3361 acggagcaag ttcccgaggt aatcggagtc cggctgatgt tgggagtagg tggctacgtc
     3421 tccgaactca cgaccgaaaa gatcaagagc agcccgcatg gatttgactt ggtcagggcc
     3481 gagcctacat gtgcgaatga tgcccatact tgagccacct aactttgttt tagggcgact
     3541 gccctgctgc gtaacatcgt tgctgctgcg taacatcgtt gctgctccat aacatcaaac
     3601 atcgacccac ggcgtaacgc gcttgctgct tggatgcccg aggcatagac tgtacaaaaa
     3661 aacagtcata acaagccatg aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca
     3721 aggttctgga ccagttgcgt gagcgcatac gctacttgca ttacagttta cgaaccgaac
     3781 aggcttatgt caactgggtt cgtgccttca tccgtttcca cggtgtgcgt cacccggcaa
     3841 ccttgggcag cagcgaagtc gaggcatttc tgtcctggct ggcgaacgag cgcaaggttt
     3901 cggtctccac gcatcgtcag gcattggcgg ccttgctgtt cttctacggc aaggtgctgt
     3961 gcacggatct gccctggctt caggagatcg gaagacctcg gccgtcgcgg cgcttgccgg
     4021 tggtgctgac cccggatgaa gtggttcgca tcctcggttt tctggaaggc gagcatcgtt
     4081 tgttcgccca ggactctagc tatagttcta gtggttggct actaataggt tgtattgatg
     4141 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg
     4201 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg
     4261 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta
     4321 attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac
     4381 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4441 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4501 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4561 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4621 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4681 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4741 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4801 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4861 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4921 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4981 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5041 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     5101 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
     5161 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
     5221 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
     5281 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
     5341 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat c
//
