LOCUS       sequence-461327-        7880 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Co-localisation experiment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7880)
  TITLE     Ori Lab colocalization and BioID plasmids 2024
REFERENCE   2  (bases 1 to 7880)
  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"
COMMENT     Sequence Label: pMX-Flag-GFP_ENPP5
FEATURES             Location/Qualifiers
     source          1..7880
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     73..92
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             266..661
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             832..1301
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(1431..1448)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1602..2190)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1682..1701)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2354..3145,3146..3214))
                     /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     2977..2996
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3215..3319)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             3349..3942
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    4005..4362
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             4421..4837
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     4767..4789
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     4805..4821
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    4847..5221
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     5159..5175
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      5250..5259
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5259..5324
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(5358..5360,5361..6071)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(5358..5360,5361..6071)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5400..5421)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5661..5680)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6008..6029
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(7559..7880,1..253,254..256,257..265)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7726..7743)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
ORIGIN
        1 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg
       61 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag
      121 gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag
      181 tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa
      241 acacgatgat aatatggcca caaccatggt tcctttgtct caagaagaat ccaccctcat
      301 tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca gcgtcgccag
      361 cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc attttactgg
      421 gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag ctggcaacct
      481 gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct gcggacggtg
      541 ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg
      601 acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt gggagggcta
      661 agtcgacgat aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc
      721 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata
      781 cataactgag aatagagaag ttcagatcaa ggtcaggaac agatggaaca gctgaatatg
      841 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
      901 gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
      961 ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc
     1021 agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca
     1081 atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc
     1141 cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt
     1201 atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt
     1261 ctcctctgag tgattgacta cccgtcagcg ggggtctttc acatgcagca tgtatcaaaa
     1321 ttaatttggt tttttttctt aagtatttac attaaatggc catagttgca ttaatgaatc
     1381 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     1441 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     1501 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     1561 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     1621 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     1681 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     1741 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     1801 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     1861 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     1921 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     1981 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2041 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2101 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2161 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2221 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2281 atcttcacct agatcctttt gcggccggcc gcaaatcaat ctaaagtata tatgagtaaa
     2341 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     2401 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     2461 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     2521 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     2581 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     2641 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     2701 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     2761 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     2821 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     2881 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     2941 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     3001 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     3061 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     3121 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     3181 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     3241 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     3301 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgcataa tgaaagaccc
     3361 cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaaatac
     3421 ataactgaga atagaaaagt tcagatcaag gtcaggaaca gatggaacag ctgaatatgg
     3481 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
     3541 aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg
     3601 gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag agaaccatca
     3661 gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa
     3721 tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc
     3781 acaacccctc actcggcgcg ccagtcctcc gattgactga gtcgcccggg tacccgtgta
     3841 tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc
     3901 tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct cgtccgggat
     3961 cgggagaccc ctgcccaggg accaccgacc caccaccggg aggtaagctg gccagcaact
     4021 tatctgtgtc tgtccgattg tctagtgtct atgactgatt ttatgcgcct gcgtcggtac
     4081 tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga gttcggaaca
     4141 cccggccgca accctgggag acgtcccagg gacttcgggg gccgtttttg tggcccgacc
     4201 tgagtccaaa aatcccgatc gttttggact ctttggtgca cccccctaat aggagggata
     4261 tgtggttctg gtaggagacg agaacctaaa acagttcccg cctccgtctg aatttttgct
     4321 ttcggtttgg gaccgaagcc gcgccgcgcg tcttgtctgc tgcagcatcg ttctgtgttg
     4381 tctctgtctg actgtgtttc tgtatttgtc tgaaaattag ggccagactg ttaccactcc
     4441 cttaagtttg accttaggtc actggaaaga tgtcgagcgg atcgctcaca accagtcggt
     4501 agatgtcaag aagagacgtt gggttacctt ctgctctgca gaatggccaa cctttaacgt
     4561 cggatggccg cgagacggca cctttaaccg agacctcatc acccaggtta agatcaaggt
     4621 cttttcacct ggcccgcatg gacacccaga ccaggtcccc tacatcgtga cctgggaagc
     4681 cttggctttt gacccccctc cctgggtcaa gccctttgta caccctaagc ctccgcctcc
     4741 tcttcctcca tccgccccgt ctctccccct tgaacctcct cgttcgaccc cgcctcgatc
     4801 ctccctttat ccagccctca ctccttctct aggcgccccc atatggccat atgagatctt
     4861 atatggggca cccccgcccc ttgtaaactt ccctgaccct gacatgacaa gagttactaa
     4921 cagcccctct ctccaagctc acttacaggc tctctactta gtccagcacg aagtctggag
     4981 acctctggcg gcagcctacc aagaacaact ggaccgaccg gtggtacctc acccttaccg
     5041 agtcggcgac acagtgtggg tccgccgaca ccagactaag aacctagaac ctcgctggaa
     5101 aggaccttac acagtcctgc tgaccacccc caccgccctc aaagtagacg gcatcgcagc
     5161 ttggatacac gccgcccacg tgaaggctgc cgaccccggg ggtggaccat cctctagact
     5221 gccggatcta gctagttaat taaggatccg ccaccatgga ctacaaggac cacgacggtg
     5281 actacaagga ccacgacatc gactacaagg acgacgacga caaggggcat ggcaccggca
     5341 gcaccggcag cgaattcgtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     5401 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     5461 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     5521 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     5581 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     5641 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     5701 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     5761 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     5821 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     5881 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     5941 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     6001 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     6061 agctgtacaa gggagggagc ggcccgcggc tcgagatgac ttcgaaattt ctcttggtgt
     6121 ccttcatact tgctgcactg agtctttcaa ccaccttttc tctccaacca gaccagcaaa
     6181 aggttctact agtttctttt gatggattcc gttgggatta cttatataaa gttccaacgc
     6241 cccattttca ttatattatg aaatatggtg ttcacgtgaa gcaagttact aatgttttta
     6301 ttacaaaaac ctaccctaac cattatactt tggtaactgg cctctttgca gagaatcatg
     6361 ggattgttgc aaatgatatg tttgatccta ttcggaacaa atctttctcc ttggatcaca
     6421 tgaatattta tgattccaag ttttgggaag aagcgacacc aatatggatc acaaaccaga
     6481 gggcaggaca tactagtggt gcagccatgt ggcccggaac agatgtaaaa atacataagc
     6541 gctttcctac tcattacatg ccttacaatg agtcagtttc atttgaagat agagttgcca
     6601 aaattattga atggtttacg tcaaaagagc ccataaatct tggtcttctc tattgggaag
     6661 accctgatga catgggccac catttgggac ctgacagtcc gctcatgggg cctgtcattt
     6721 cagatattga caagaagtta ggatatctca tacaaatgct gaaaaaggca aagttgtgga
     6781 acactctgaa cctaatcatc acaagtgatc atggaatgac gcagtgctct gaggaaaggt
     6841 taatagaact tgaccagtac ctggataaag accactatac cctgattgat caatctccag
     6901 tagcagccat cttgccaaaa gaaggtaaat ttgatgaagt ctatgaagca ctaactcacg
     6961 ctcatcctaa tcttactgtt tacaaaaaag aagacgttcc agaaaggtgg cattacaaat
     7021 acaacagtcg aattcaacca atcatagcag tggctgatga agggtggcac attttacaga
     7081 ataagtcaga tgactttctg ttaggcaacc acggttacga taatgcgtta gcagatatgc
     7141 atccaatatt tttagcccat ggtcctgcct tcagaaagaa tttctcaaaa gaagccatga
     7201 actccacaga tttgtaccca ctactatgcc acctcctcaa tatcaccgcc atgccacaca
     7261 atggatcatt ctggaatgtc caggatctgc tcaattcagc aatgccaagg gtggtccctt
     7321 atacacagag tactatactc ctccctggta gtgttaaacc agcagaatat gaccaagagg
     7381 ggtcataccc ttatttcata ggggtctctc ttggcagcat tatagtgatt gtattttttg
     7441 taattttcat taagcattta attcacagtc aaatacctgc cttacaagat atgcatgctg
     7501 aaatagctca accattatta caagcctaag cggccgctac gtaaattccg cccccccccc
     7561 cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg
     7621 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga
     7681 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa
     7741 tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa
     7801 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct
     7861 gcggccaaaa gccacgtgta
//
