LOCUS       sequence-420367-        8795 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of untagged MinE in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8795)
  AUTHORS   Rajasekaran R, Chang CC, Weix EWZ, Galateo TM, Coyle SM
  TITLE     A programmable reaction-diffusion system for spatiotemporal cell 
            signaling circuit design.
  JOURNAL   Cell. 2023 Dec 27:S0092-8674(23)01339-9. doi: 
            10.1016/j.cell.2023.12.007.
  PUBMED    38181787
REFERENCE   2  (bases 1 to 8795)
  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: "Cell. 2023 
            Dec 27:S0092-8674(23)01339-9. doi: 10.1016/j.cell.2023.12.007."
COMMENT     Sequence Label: pLV-EF1a-MinE-IRES-Blast
FEATURES             Location/Qualifiers
     source          1..8795
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(202..213)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             526..1159
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(1287..1303)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1287..1303)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1300..1322)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1311..1327
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1335..1341,1342..1359,1360..1365))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1380..1401
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1518..1535)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1689..2277)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1769..1788)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2448..3239,3240..3308))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3071..3090
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3309..3413)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    3461..3595
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3498..3517)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3552..3571
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             3763..4396
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    4443..4568
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5065..5298
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5483..5527
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    5790..5905
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        5983..6194
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          6195..7133
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     7090..7110
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      7149..7158
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    7522..8071
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7666..7683)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7893..7912
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     misc_feature    join(8526..8795,1..319)
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8579..8599)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
ORIGIN
        1 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
       61 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
      121 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
      181 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
      241 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
      301 ctttgggccg cctccccgcc tggaattaat tctgcagtcg agacctagaa aaacatggag
      361 caatcacaag tagcaataca gcagctacca atgctgattg tgcctggcta gaagcacaag
      421 aggaggagga ggtgggtttt ccagtcacac ctcaggtacc tttaagacca atgacttaca
      481 aggcagctgt agatcttagc cactttttaa aagaaaagag gggactggaa gggctaattc
      541 actcccaacg aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc
      601 ctgattagca gaactacaca ccagggccag gggtcagata tccactgacc tttggatggt
      661 gctacaagct agtaccagtt gagccagata aggtagaaga ggccaataaa ggagagaaca
      721 ccagcttgtt acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtgttag
      781 agtggaggtt tgacagccgc ctagcatttc atcacgtggc ccgagagctg catccggagt
      841 acttcaagaa ctgctgatat cgagcttgct acaagggact ttccgctggg gactttccag
      901 ggaggcgtgg cctgggcggg actggggagt ggcgagccct cagatcctgc atataagcag
      961 ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     1021 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     1081 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     1141 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt
     1201 gcaaagaaat gaatatcaga gagtgagagg ccttgacatt gctagcgttt accgtcgacc
     1261 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     1321 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     1381 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     1441 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     1501 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     1561 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     1621 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     1681 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     1741 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     1801 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     1861 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     1921 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     1981 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2041 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2101 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     2161 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2221 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2281 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     2341 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     2401 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     2461 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     2521 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     2581 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     2641 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     2701 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     2761 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     2821 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     2881 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     2941 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3001 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3061 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     3121 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3181 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     3241 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3301 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     3361 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     3421 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc aacttgttta ttgcagctta
     3481 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3541 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaactg
     3601 gataactcaa gctaaccaaa atcatcccaa acttcccacc ccatacccta ttaccactgc
     3661 caattacctg tggtttcatt tactctaaac ctgtgattcc tctgaattat tttcatttta
     3721 aagaaattgt atttgttaaa tatgtactac aaacttagta gttggaaggg ctaattcact
     3781 cccaaagaag acaagatatc cttgatctgt ggatctacca cacacaaggc tacttccctg
     3841 attagcagaa ctacacacca gggccagggg tcagatatcc actgaccttt ggatggtgct
     3901 acaagctagt accagttgag ccagataagg tagaagaggc caataaagga gagaacacca
     3961 gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa gtgttagagt
     4021 ggaggtttga cagccgccta gcatttcatc acgtggcccg agagctgcat ccggagtact
     4081 tcaagaactg ctgatatcga gcttgctaca agggactttc cgctggggac tttccaggga
     4141 ggcgtggcct gggcgggact ggggagtggc gagccctcag atcctgcata taagcagctg
     4201 ctttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     4261 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     4321 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     4381 tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg
     4441 agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc
     4501 gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg
     4561 agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc
     4621 cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac
     4681 gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac
     4741 agctacaacc atcccttcag acaggatcag aagaacttag atcattatat aatacagtag
     4801 caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca
     4861 agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc ggccgctgat
     4921 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
     4981 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
     5041 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
     5101 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
     5161 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
     5221 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
     5281 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
     5341 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
     5401 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
     5461 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
     5521 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
     5581 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
     5641 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
     5701 cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg
     5761 attagtgaac ggatctcgac ggtatcgcct ttaaaagaaa aggggggatt ggggggtaca
     5821 gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa
     5881 aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt
     5941 atcgatgagg ccctttcgtc ttcactcgag gtgcccgtca gtgggcagag cgcacatcgc
     6001 ccacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc tagagaaggt
     6061 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
     6121 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
     6181 ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt
     6241 atggcccttg cgtgccttga attacttcca cctggctgca gtacgtgatt cttgatcccg
     6301 agcttcgggt tggaagtggg tgggagagtt cgaggccttg cgcttaagga gccccttcgc
     6361 ctcgtgcttg agttgaggcc tggcctgggc gctggggccg ccgcgtgcga atctggtggc
     6421 accttcgcgc ctgtctcgct gctttcgata agtctctagc catttaaaat ttttgatgac
     6481 ctgctgcgac gctttttttc tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca
     6541 ctggtatttc ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat
     6601 gttcggcgag gcggggcctg cgagcgcggc caccgagaat cggacggggg tagtctcaag
     6661 ctggccggcc tgctctggtg cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg
     6721 caaggctggc ccggtcggca ccagttgcgt gagcggaaag atggccgctt cccggccctg
     6781 ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt gagtcaccca
     6841 cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc
     6901 gggcgccgtc caggcacctc gattagttct cgagcttttg gagtacgtcg tctttaggtt
     6961 ggggggaggg gttttatgcg atggagtttc cccacactga gtgggtggag actgaagtta
     7021 ggccagcttg gcacttgatg taattctcct tggaatttgc cctttttgag tttggatctt
     7081 ggttcattct caagcctcag acagtggttc aaagtttttt tcttccattt caggtgtcgt
     7141 gaggatccgc caccatggcc cttcttgact tcttcctgtc tcgaaaaaag aacacagcga
     7201 atatcgcgaa agaacggttg caaattatag tggcggagag acgacgaagc gacgcggagc
     7261 ctcattacct tcctcaattg cgaaaagaca tactggaagt gatctgcaag tacgttcaaa
     7321 ttgaccctga gatggtgacc gtgcaacttg aacagaagga tggagatata agcatcctgg
     7381 aattgaacgt aacccttccc gaggcggaag aacttaaata agcggccgct ctagagtcga
     7441 cgggccgcgg taacaattgt taactaactt aagctagcaa cggtttccct ctagcgggat
     7501 caattccgcc ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg
     7561 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga
     7621 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa
     7681 tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa
     7741 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct
     7801 gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg
     7861 ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg
     7921 ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca
     7981 catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg
     8041 acgtggtttt cctttgaaaa acacgataat accatggtca tgaaaacatt taacatttct
     8101 caacaagatc tagaattagt agaagtagcg acagagaaga ttacaatgct ttatgaggat
     8161 aataaacatc atgtgggagc ggcaattcgt acgaaaacag gagaaatcat ttcggcagta
     8221 catattgaag cgtatatagg acgagtaact gtttgtgcag aagccattgc gattggtagt
     8281 gcagtttcga atggacaaaa ggattttgac acgattgtag ctgttagaca cccttattct
     8341 gacgaagtag atagaagtat tcgagtggta agtccttgtg gtatgtgtag ggagttgatt
     8401 tcagactatg caccagattg ttttgtgtta atagaaatga atggcaagtt agtcaaaact
     8461 acgattgaag aactcattcc actcaaatat acccgaaatt aacccgggcg gggcgcgtct
     8521 ggaacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     8581 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     8641 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     8701 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     8761 ccactggttg gggcattgcc accacctgtc agctc
//
