LOCUS       sequence-461332-        7889 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 7889)
  TITLE     Ori Lab colocalization and BioID plasmids 2024
REFERENCE   2  (bases 1 to 7889)
  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_EDIL3
FEATURES             Location/Qualifiers
     source          1..7889
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..40
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(259..363)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             393..986
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    1049..1406
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1465..1881
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1811..1833
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1849..1865
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1891..2265
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     2203..2219
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      2294..2303
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2303..2368
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(2402..2404,2405..3115)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2402..2404,2405..3115)
                     /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(2444..2465)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2705..2724)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3052..3073
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(4612..5186,5187..5189,5190..5198)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4779..4796)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5006..5025
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5199..5594
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             5765..6234
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6364..6381)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6535..7123)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6615..6634)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7287..7889,1..189,190..258))
                     /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"
ORIGIN
        1 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
       61 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
      121 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
      181 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
      241 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
      301 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
      361 gcgcacattt ccccgaaaag tgccacctgc ataatgaaag accccacctg taggtttggc
      421 aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagaa
      481 aagttcagat caaggtcagg aacagatgga acagctgaat atgggccaaa caggatatct
      541 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggaacagc tgaatatggg
      601 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt
      661 ccccagatgc ggtccagccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc
      721 cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc
      781 ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg
      841 cgcgccagtc ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt
      901 gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga
      961 ctacccgtca gcgggggtct ttcatttggg ggctcgtccg ggatcgggag acccctgccc
     1021 agggaccacc gacccaccac cgggaggtaa gctggccagc aacttatctg tgtctgtccg
     1081 attgtctagt gtctatgact gattttatgc gcctgcgtcg gtactagtta gctaactagc
     1141 tctgtatctg gcggacccgt ggtggaactg acgagttcgg aacacccggc cgcaaccctg
     1201 ggagacgtcc cagggacttc gggggccgtt tttgtggccc gacctgagtc caaaaatccc
     1261 gatcgttttg gactctttgg tgcacccccc taataggagg gatatgtggt tctggtagga
     1321 gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttgggaccga
     1381 agccgcgccg cgcgtcttgt ctgctgcagc atcgttctgt gttgtctctg tctgactgtg
     1441 tttctgtatt tgtctgaaaa ttagggccag actgttacca ctcccttaag tttgacctta
     1501 ggtcactgga aagatgtcga gcggatcgct cacaaccagt cggtagatgt caagaagaga
     1561 cgttgggtta ccttctgctc tgcagaatgg ccaaccttta acgtcggatg gccgcgagac
     1621 ggcaccttta accgagacct catcacccag gttaagatca aggtcttttc acctggcccg
     1681 catggacacc cagaccaggt cccctacatc gtgacctggg aagccttggc ttttgacccc
     1741 cctccctggg tcaagccctt tgtacaccct aagcctccgc ctcctcttcc tccatccgcc
     1801 ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct ttatccagcc
     1861 ctcactcctt ctctaggcgc ccccatatgg ccatatgaga tcttatatgg ggcacccccg
     1921 ccccttgtaa acttccctga ccctgacatg acaagagtta ctaacagccc ctctctccaa
     1981 gctcacttac aggctctcta cttagtccag cacgaagtct ggagacctct ggcggcagcc
     2041 taccaagaac aactggaccg accggtggta cctcaccctt accgagtcgg cgacacagtg
     2101 tgggtccgcc gacaccagac taagaaccta gaacctcgct ggaaaggacc ttacacagtc
     2161 ctgctgacca cccccaccgc cctcaaagta gacggcatcg cagcttggat acacgccgcc
     2221 cacgtgaagg ctgccgaccc cgggggtgga ccatcctcta gactgccgga tctagctagt
     2281 taattaagga tccgccacca tggactacaa ggaccacgac ggtgactaca aggaccacga
     2341 catcgactac aaggacgacg acgacaaggg gcatggcacc ggcagcaccg gcagcgaatt
     2401 cgtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     2461 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     2521 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     2581 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     2641 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     2701 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     2761 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     2821 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     2881 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     2941 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     3001 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     3061 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggagg
     3121 gagcggcccg cggctcgaga tgaagcgctc ggtagccgtc tggctcttgg tcgggctcag
     3181 cctcggtgtc ccccagttcg gcaaaggtga tatttgtgat cccaatccat gtgaaaatgg
     3241 aggtatctgt ttgccaggat tggctgatgg ttccttttcc tgtgagtgtc cagatggctt
     3301 cacagacccc aactgttcta gtgttgtgga ggttgcatca gatgaagaag aaccaacttc
     3361 agcaggtccc tgcactccta atccatgcca taatggagga acctgtgaaa taagtgaagc
     3421 ataccgaggg gatacattca taggctatgt ttgtaaatgt ccccgaggat ttaatgggat
     3481 tcactgtcag cacaacataa atgaatgcga agttgagcct tgcaaaaatg gtggaatatg
     3541 tacagatctt gttgctaact attcctgtga gtgcccaggc gaatttatgg gaagaaattg
     3601 tcaatacaaa tgctcaggcc cactgggaat tgaaggtgga attatatcaa accagcaaat
     3661 cacagcttcc tctactcacc gagctctttt tggactccaa aaatggtatc cctactatgc
     3721 acgtcttaat aagaaggggc ttataaatgc gtggacagct gcagaaaatg acagatggcc
     3781 gtggattcag ataaatttgc aaaggaaaat gagagttact ggtgtgatta cccaaggagc
     3841 caagaggatt ggaagcccag agtatataaa atcctacaaa attgcctaca gtaatgatgg
     3901 aaagacttgg gcaatgtaca aagtgaaagg caccaatgaa gacatggtgt ttcgtggaaa
     3961 cattgataac aacactccat atgctaactc tttcacaccc cccataaaag ctcagtatgt
     4021 aagactctat ccccaagttt gtcgaagaca ttgcactttg cgaatggaac ttcttggctg
     4081 tgaactgtcg ggttgttctg agcctctggg tatgaaatca ggacatatac aagactatca
     4141 gatcactgcc tccagcatct tcagaacgct caacatggac atgttcactt gggaaccaag
     4201 gaaagctcgg ctggacaagc aaggcaaagt gaatgcctgg acctctggcc acaatgacca
     4261 gtcacaatgg ttacaggtgg atcttcttgt tccaaccaaa gtgactggca tcattacaca
     4321 aggagctaaa gattttggtc atgtacagtt tgttggctcc tacaaactgg cttacagcaa
     4381 tgatggagaa cactggactg tataccagga tgaaaagcaa agaaaagata aggttttcca
     4441 gggaaatttt gacaatgaca ctcacagaaa aaatgtcatc gaccctccca tctatgcacg
     4501 acacataaga atccttcctt ggtcctggta cgggaggatc acattgcggt cagagctgct
     4561 gggctgcaca gaggaggaat gagcggccgc tacgtaaatt ccgccccccc ccccctctcc
     4621 ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt
     4681 ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg
     4741 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg
     4801 tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc
     4861 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca
     4921 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag
     4981 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa
     5041 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt
     5101 tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt
     5161 tttcctttga aaaacacgat gataatatgg ccacaaccat ggttcctttg tctcaagaag
     5221 aatccaccct cattgaaaga gcaacggcta caatcaacag catccccatc tctgaagact
     5281 acagcgtcgc cagcgcagct ctctctagcg acggccgcat cttcactggt gtcaatgtat
     5341 atcattttac tgggggacct tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg
     5401 cagctggcaa cctgacttgt atcgtcgcga tcggaaatga gaacaggggc atcttgagcc
     5461 cctgcggacg gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa gccatagtga
     5521 aggacagtga tggacagccg acggcagttg ggattcgtga attgctgccc tctggttatg
     5581 tgtgggaggg ctaagtcgac gataaaataa aagattttat ttagtctcca gaaaaagggg
     5641 ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag
     5701 gcatggaaaa atacataact gagaatagag aagttcagat caaggtcagg aacagatgga
     5761 acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     5821 ccaagaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc
     5881 tgccccggct cagggccaag aacagatggt ccccagatgc ggtccagccc tcagcagttt
     5941 ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta
     6001 tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct
     6061 caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc
     6121 cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt
     6181 tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcacatgca
     6241 gcatgtatca aaattaattt ggtttttttt cttaagtatt tacattaaat ggccatagtt
     6301 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     6361 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     6421 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6481 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6541 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6601 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6661 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6721 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6781 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6841 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6901 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6961 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7021 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     7081 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     7141 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     7201 attatcaaaa aggatcttca cctagatcct tttgcggccg gccgcaaatc aatctaaagt
     7261 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7321 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7381 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7441 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7501 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7561 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7621 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7681 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7741 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7801 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7861 gaatagtgta tgcggcgacc gagttgctc
//
