LOCUS       sequence-455397-        7172 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 7172)
  TITLE     Ori Lab colocalization and BioID plasmids 2024
REFERENCE   2  (bases 1 to 7172)
  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_PDGFA
FEATURES             Location/Qualifiers
     source          1..7172
                     /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(3895..4469,4470..4472,4473..4481)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4062..4079)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4289..4308
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4482..4877
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             5048..5517
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5647..5664)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5818..6406)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5898..5917)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6570..7172,1..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 tgtgggtgga gggtcctcat tcagatgccc caggcctcgg
     3181 atcctgggcg ggggcgcgtg gcagccgcca ggagaccggc tggagcgccc gccccgcggc
     3241 ctcgcctctc ctccgagcag ccagcgcctc gggacgcgat gaggaccttg gcttgcctgc
     3301 tgctcctcgg ctgcggatac ctcgcccatg ttctggccga ggaagccgag atcccccgcg
     3361 aggtgatcga gaggctggcc cgcagtcaga tccacagcat ccgggacctc cagcgactcc
     3421 tggagataga ctccggagtg aggattcttt ggacaccagc ctgagagctc acggggtcca
     3481 tgccactaag catgtgcccg agaagcggcc cctgcccatt cggaggaaga gaagcatcga
     3541 ggaagctgtc cccgctgtct gcaagaccag gacggtcatt tacgagattc ctcggagtca
     3601 ggtcgacccc acgtccgcca acttcctgat ctggcccccg tgcgtggagg tgaaacgctg
     3661 caccggctgc tgcaacacga gcagtgtcaa gtgccagccc tcccgcgtcc accaccgcag
     3721 cgtcaaggtg gccaaggtgg aatacgtcag gaagaagcca aaattaaaag aagtccaggt
     3781 gaggttagag gagcatttgg agtgcgcctg cgcgaccaca agcctgaatc cggattatcg
     3841 ggaagaggac acggatgtga ggtgagcggc cgctacgtaa attccgcccc cccccccctc
     3901 tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt
     3961 tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc
     4021 tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca
     4081 aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac
     4141 gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg
     4201 ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt
     4261 gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct
     4321 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg
     4381 ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt
     4441 ggttttcctt tgaaaaacac gatgataata tggccacaac catggttcct ttgtctcaag
     4501 aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc atctctgaag
     4561 actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact ggtgtcaatg
     4621 tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact gctgctgctg
     4681 cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga
     4741 gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc aaagccatag
     4801 tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg ccctctggtt
     4861 atgtgtggga gggctaagtc gacgataaaa taaaagattt tatttagtct ccagaaaaag
     4921 gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc
     4981 aaggcatgga aaaatacata actgagaata gagaagttca gatcaaggtc aggaacagat
     5041 ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
     5101 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
     5161 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
     5221 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
     5281 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
     5341 gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat tgactgagtc
     5401 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
     5461 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcacat
     5521 gcagcatgta tcaaaattaa tttggttttt tttcttaagt atttacatta aatggccata
     5581 gttgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5641 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5701 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5761 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5821 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5881 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5941 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6001 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6061 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6121 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6181 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6241 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6301 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6361 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6421 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6481 gagattatca aaaaggatct tcacctagat ccttttgcgg ccggccgcaa atcaatctaa
     6541 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6601 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6661 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6721 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6781 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6841 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6901 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6961 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7021 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7081 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7141 tgagaatagt gtatgcggcg accgagttgc tc
//
