LOCUS       sequence-436554-        8058 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral mammalian expression vector with GFP for overexpression 
            of mouse PHGDH.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8058)
  TITLE     Jones lab mouse PHGDH plasmid
REFERENCE   2  (bases 1 to 8058)
  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: MigR1 GFP Mouse PHGDH
FEATURES             Location/Qualifiers
     source          1..8058
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             12..526
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    688..704
                     /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(712..718,719..736,737..742))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    757..778
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(895..912)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1066..1654)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1146..1165)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1825..2616,2617..2685))
                     /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     2448..2467
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2686..2790)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2858..2876
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2914..2936)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3036..3055
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3249..3271
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3399..3418
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             3618..4134
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    4198..4539
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             4606..5022
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     4952..4974
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     4990..5006
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      5049..5058
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    6668..7235
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    6685..7237
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6829..6846)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7056..7075
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(7249..7251,7252..7254,7255..7968)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7294..7315)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7555..7574)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7902..7923
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat
       61 tttgcaaggc atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac
      121 agagagacag cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc
      181 tcagggccaa gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac
      241 catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta
      301 accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag
      361 agcccacaac ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc
      421 gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga
      481 gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatgggt aacagtttct
      541 tgaagttgga gaacaacatt ctgagggtag gagtcgaata ttaagtaatc ctgactcaat
      601 tagccactgt tttgaatcca catactccaa tactcctgaa atagttcatt atggacagcg
      661 cagaaagagc tggggagaat tgtgaaattg ttatccgctc acaattccac acaacatacg
      721 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
      781 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
      841 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
      901 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
      961 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     1021 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     1081 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1141 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1201 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1261 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     1321 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     1381 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     1441 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     1501 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     1561 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     1621 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     1681 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     1741 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     1801 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     1861 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     1921 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     1981 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     2041 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     2101 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     2161 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     2221 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     2281 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     2341 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     2401 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     2461 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     2521 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     2581 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     2641 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     2701 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     2761 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     2821 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     2881 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     2941 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     3001 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     3061 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     3121 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     3181 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     3241 agggttttcc cagtcacgac gttgtaaaac gacggcgcaa ggaatggtgc atgcaaggag
     3301 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
     3361 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg
     3421 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggcga
     3481 ttagtccaat ttgttaaaga caggatatca gtggtccagg ctctagtttt gactcaacaa
     3541 tatcaccagc tgaagcctat agagtacgag ccatagataa aataaaagat tttatttagt
     3601 ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta
     3661 acgccatttt gcaaggcatg gaaaatacat aactgagaat agagaagttc agatcaaggt
     3721 taggaacaga gagacagcag aatatgggcc aaacaggata tctgtggtaa gcagttcctg
     3781 ccccggctca gggccaagaa cagatggtcc ccagatgcgg tcccgccctc agcagtttct
     3841 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
     3901 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
     3961 ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgatagact gcgtcgcccg
     4021 ggtacccgta ttcccaataa agcctcttgc tgtttgcatc cgaatcgtgg actcgctgat
     4081 ccttgggagg gtctcctcag attgattgac tgcccacctc gggggtcttt catttggagg
     4141 ttccaccgag atttggagac ccctgcccag ggaccaccga cccccccgcc gggaggtaag
     4201 ctggccagcg gtcgtttcgt gtctgtctct gtctttgtgc gtgtttgtgc cggcatctaa
     4261 tgtttgcgcc tgcgtctgta ctagttagct aactagctct gtatctggcg gacccgtggt
     4321 ggaactgacg agttctgaac acccggccgc aaccctggga gacgtcccag ggactttggg
     4381 ggccgttttt gtggcccgac ctgaggaagg gagtcgatgt ggaatccgac cccgtcagga
     4441 tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc tgaatttttg
     4501 ctttcggttt ggaaccgaag ccgcgcgtct tgtctgctgc agcgctgcag catcgttctg
     4561 tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa attagggcca gactgttacc
     4621 actcccttaa gtttgacctt aggtcactgg aaagatgtcg agcggatcgc tcacaaccag
     4681 tcggtagatg tcaagaagag acgttgggtt accttctgct ctgcagaatg gccaaccttt
     4741 aacgtcggat ggccgcgaga cggcaccttt aaccgagacc tcatcaccca ggttaagatc
     4801 aaggtctttt cacctggccc gcatggacac ccagaccagg tcccctacat cgtgacctgg
     4861 gaagccttgg cttttgaccc ccctccctgg gtcaagccct ttgtacaccc taagcctccg
     4921 cctcctcttc ctccatccgc cccgtctctc ccccttgaac ctcctcgttc gaccccgcct
     4981 cgatcctccc tttatccagc cctcactcct tctctaggcg ccggaattag atctctcgag
     5041 gttagcttgc caccatggct tttgcaaatc tgcggaaagt gctcatcagt gacagcctgg
     5101 acccttgctg ccggaagatc ttgcaagatg gagggctgca ggtggtggaa aagcagaacc
     5161 ttagcaaaga ggagctgata gcggagctgc aggactgtga aggccttatt gttcgctctg
     5221 ccaccaaggt gaccgctgat gtcatcaacg cagctgagaa actccaggtg gtgggcaggg
     5281 ctggcacagg tgtggacaat gtggatctgg aggccgcaac aaggaagggc atcttggtta
     5341 tgaacacccc caatgggaac agcctcagtg ccgcagaact cacttgtgga atgatcatgt
     5401 gcctggccag gcagattccc caggcgacgg cttcgatgaa ggacggcaaa tgggagcgga
     5461 agaagttcat gggaacagag ctgaatggaa agaccctggg aattcttggc ctgggcagga
     5521 ttgggagaga ggtagctacc cggatgcagt cctttgggat gaagactata gggtatgacc
     5581 ccatcatttc cccagaggtc tcggcctcct ttggtgttca gcagctgccc ctggaggaga
     5641 tctggcctct ctgtgatttc atcactgtgc acactcctct cctgccctcc acgacaggct
     5701 tgctgaatga caacaccttt gcccagtgca agaagggggt gcgtgtggtg aactgtgccc
     5761 gtggagggat cgtggacgaa ggcgccctgc tccgggccct gcagtctggc cagtgtgccg
     5821 gggctgcact ggacgtgttt acggaagagc cgccacggga ccgggccttg gtggaccatg
     5881 agaatgtcat cagctgtccc cacctgggtg ccagcaccaa ggaggctcag agccgctgtg
     5941 gggaggaaat tgctgttcag ttcgtggaca tggtgaaggg gaaatctctc acgggggttg
     6001 tgaatgccca ggcccttacc agtgccttct ctccacacac caagccttgg attggtctgg
     6061 cagaagctct ggggacactg atgcgagcct gggctgggtc ccccaaaggg accatccagg
     6121 tgataacaca gggaacatcc ctgaagaatg ctgggaactg cctaagcccc gcagtcattg
     6181 tcggcctcct gaaagaggct tccaagcagg cggatgtgaa cttggtgaac gctaagctgc
     6241 tggtgaaaga ggctggcctc aatgtcacca cctcccacag ccctgctgca ccaggggagc
     6301 aaggcttcgg ggaatgcctc ctggccgtgg ccctggcagg cgccccttac caggctgtgg
     6361 gcttggtcca aggcactaca cctgtactgc aggggctcaa tggagctgtc ttcaggccag
     6421 aagtgcctct ccgcagggac ctgcccctgc tcctattccg gactcagacc tctgaccctg
     6481 caatgctgcc taccatgatt ggcctcctgg cagaggcagg cgtgcggctg ctgtcctacc
     6541 agacttcact ggtgtcagat ggggagacct ggcacgtcat gggcatctcc tccttgctgc
     6601 ccagcctgga agcgtggaag cagcatgtga ctgaagcctt ccagttccac ttctaaatcg
     6661 aacgaattcc gccccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt
     6721 gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc
     6781 ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag
     6841 gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac
     6901 aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc
     6961 tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc
     7021 acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca
     7081 aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt
     7141 gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg
     7201 gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat ggtgagcaag
     7261 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     7321 ggccacaagt tcagcgtgtc tggcgagggc gagggcgatg ccacctacgg caagctgacc
     7381 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     7441 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     7501 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     7561 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     7621 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     7681 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggcg
     7741 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     7801 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     7861 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     7921 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaatg aattaattaa
     7981 gaattatcaa gcttatcgat accgtcgacc tgcagccaag cttatcgata aaataaaaga
     8041 ttttatttag tctccaga
//
