LOCUS       sequence-455405-        8795 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 8795)
  TITLE     Ori Lab colocalization and BioID plasmids 2024
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"
COMMENT     Sequence Label: pMX-Flag-GFP_SLC45A1
FEATURES             Location/Qualifiers
     source          1..8795
                     /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(5518..6092,6093..6095,6096..6104)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5685..5702)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5912..5931
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6105..6500
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             6671..7140
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(7270..7287)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7441..8029)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7521..7540)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8193..8795,1..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 tgctgcagca gccaggacct aggccaggcc gccagcagcc
     3181 ctccggcgac agggatgcat gcaggctgca ccctcagggc aggccccctg ccctgccaac
     3241 catgatccct gcagcaagct ccacaccacc aggcgacgca ctgttcccaa gcgtggcacc
     3301 ccaggatttt tggcgctccc aggtgaccgg atacagcgga tccgtgacaa ggcacctgtc
     3361 ccaccgcgcc aacaacttca agcggcaccc taagcggaga aagtgcatca gaccaagccc
     3421 tccaccccct ccaaacaccc cctgtcctct ggagctggtg gacttcggcg atctgcaccc
     3481 ccagcggtcc ttcagagagc tgctgtttaa tggctgcatc ctgttcggca tcgagttttc
     3541 ttacgccatg gagaccgcct atgtgacacc cgtgctgctg cagatgggcc tgcctgacca
     3601 gctgtatagc ctggtgtggt tcatctcccc aatcctgggc tttctgctgc agccactgct
     3661 gggagcatgg tctgataggt gtaccagcag attcggcagg cgccggccct ttatcctggt
     3721 gctggcaatc ggcgccctgc tgggactgag cctgctgctg aacggaaggg acatcggaat
     3781 cgccctggca gatgtgaccg gcaatcacaa gtggggactg ctgctgacag tgtgcggagt
     3841 ggtgctgatg gacttcagcg ccgactccgc cgataaccca tctcacgcct acatgatgga
     3901 cgtgtgcagc cccgccgacc aggatagagg cctgaatatc cacgcactgc tggcaggact
     3961 gggaggaggc ttcggctacg tggtgggcgg catccactgg gataagaccg gatttggacg
     4021 ggccctggga ggacagctga gagtgatcta tctgttcacc gccgtgacac tgtccgtgac
     4081 cacagtgctg acactggtgt ctatcccaga gaggccactg aggccacctt ctgagaagag
     4141 ggccgccatg aagtctccta gcctgccact gccaccaagc cctccagtgc tgcctgagga
     4201 gggaccaggc gactctctgc ctagccacac cgccacaaac ttttctagcc caatctcccc
     4261 cccttctccc ctgaccccta agtacggcag cttcatctcc cgcgattcct ctctgaccgg
     4321 catcagcgag ttcgccagct cctttggcac agccaatatc gactccgtgc tgatcgattg
     4381 ctttacagga ggacacgact cctacctggc aatcccagga tctgtgccta ggccacccat
     4441 ctccgtgtct ttcccaaggg caccagacgg cttttatcgg caggatagag gactgctgga
     4501 gggaagggag ggcgccctga ccagcggctg tgacggcgat atcctgcgcg tgggctccct
     4561 ggacacctct aagcccaggt ctagcggcat cctgaagcgc ccacagacac tggcaatccc
     4621 agatgcagca ggaggaggag gccctgagac cagcagaagg cgcaacgtga cattctccca
     4681 gcaggtggcc aacatcctgc tgaatggcgt gaagtacgag tccgagctga ccggatcctc
     4741 tgagagggca gagcagcctc tgtctgtggg cagactgtgc agcaccatct gtaacatgcc
     4801 aaaggccctg aggacactgt gcgtgaatca cttcctgggc tggctgagct ttgagggcat
     4861 gctgctgttc tataccgact ttatgggcga ggtggtgttt cagggcgatc ctaaggcccc
     4921 acacacctct gaggcctacc agaagtataa cagcggcgtg acaatgggct gctggggcat
     4981 gtgcatctac gccttctccg ccgcctttta ttctgccatc ctggagaagc tggaggagtt
     5041 cctgtccgtg cggaccctgt acttcatcgc ctatctggcc tttggcctgg gaaccggact
     5101 ggccacactg tccagaaatc tgtacgtggt gctgtctctg tgcatcacat atggcatcct
     5161 gtttagcacc ctgtgcacac tgccctactc cctgctgtgc gactactatc agtctaagaa
     5221 gttcgcagga agctccgccg acggaaccag gagaggcatg ggcgtggata tctctctgct
     5281 gagctgccag tactttctgg cccagatcct ggtgagcctg gtgctgggac cactgacaag
     5341 cgccgtggga tccgccaacg gcgtgatgta tttctctagc ctggtgtcct ttctgggctg
     5401 tctgtactcc tctctgttcg tgatctatga gatccctcca agcgacgccg ccgatgagga
     5461 gcacagacct ctgctgctga atgtgtgagc ggccgctacg taaattccgc cccccccccc
     5521 ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc
     5581 gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa
     5641 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat
     5701 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac
     5761 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg
     5821 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt
     5881 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg
     5941 gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac
     6001 atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga
     6061 cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatggtt cctttgtctc
     6121 aagaagaatc caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg
     6181 aagactacag cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca
     6241 atgtatatca ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg
     6301 ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct
     6361 tgagcccctg cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca
     6421 tagtgaagga cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg
     6481 gttatgtgtg ggagggctaa gtcgacgata aaataaaaga ttttatttag tctccagaaa
     6541 aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt
     6601 tgcaaggcat ggaaaaatac ataactgaga atagagaagt tcagatcaag gtcaggaaca
     6661 gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc
     6721 tcagggccaa gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc
     6781 agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag
     6841 cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt
     6901 gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc
     6961 cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc gattgactga
     7021 gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct
     7081 cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca
     7141 catgcagcat gtatcaaaat taatttggtt ttttttctta agtatttaca ttaaatggcc
     7201 atagttgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     7261 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     7321 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     7381 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     7441 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     7501 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     7561 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     7621 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     7681 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7741 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7801 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7861 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     7921 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7981 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     8041 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     8101 catgagatta tcaaaaagga tcttcaccta gatccttttg cggccggccg caaatcaatc
     8161 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     8221 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     8281 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     8341 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     8401 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     8461 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     8521 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     8581 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     8641 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     8701 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     8761 ttctgagaat agtgtatgcg gcgaccgagt tgctc
//
