LOCUS       sequence-213568-        6899 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian retroviral expression of glutaminase isoform KGA (S286A) 
            with V5 tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6899)
  TITLE     Mammalian retroviral expression plasmids based on pBABE family
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6899)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pBABE.zeo/hKGA.S286A
FEATURES             Location/Qualifiers
     source          1..6899
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..470
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    535..734
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             935..1351
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1281..1303
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1319..1335
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             3441..3482
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3497..3517)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3502..3831
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3682..3817
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3841..4215
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     LTR             4386..4855
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4985..5002)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5156..5744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5236..5255)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5904..6695,6696..6764))
                     /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     6527..6546
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6765..6869)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
       61 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
      121 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
      181 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
      241 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
      301 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
      361 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
      421 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct
      481 cgtccgggat cgggagaccc ctgcccaggg accaccgacc caccaccggg aggcaagctg
      541 gccagcaact tatctgtgtc tgtccgattg tctagtgtct atgactgatt ttatgcgcct
      601 gcgtcggtac tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga
      661 gttctgaaca cccggccgca accctgggag acgtcccagg gactttgggg gccgtttttg
      721 tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat atgtggttct
      781 ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg
      841 gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt gttgtctctg
      901 tctgactgtg tttctgtatt tgtctgaaaa ttagggccag actgttacca ctcccttaag
      961 tttgacctta ggtcactgga aagatgtcga gcggatcgct cacaaccagt cggtagatgt
     1021 caagaagaga cgttgggtta ccttctgctc tgcagaatgg ccaaccttta acgtcggatg
     1081 gccgcgagac ggcaccttta accgagacct catcacccag gttaagatca aggtcttttc
     1141 acctggcccg catggacacc cagaccaggt cccctacatc gtgacctggg aagccttggc
     1201 ttttgacccc cctccctggg tcaagccctt tgtacaccct aagcctccgc ctcctcttcc
     1261 tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct
     1321 ttatccagcc ctcactcctt ctctaggcgc cggccggatc catgatgcgg ctgcgaggct
     1381 cggggatgct gcgggacctg ctcctgcggt cgcccgccgg cgtgagcgcg actctgcggc
     1441 gggcacagcc cttggtcacc ctgtgccggc gtccccgagg cgggggacgg ccggccgcgg
     1501 gcccggctgc cgccgcgcga ctccacccgt ggtggggcgg gggcggctgg ccggcggagc
     1561 ccctcgcgcg gggcctgtcc agctctcctt cggagatctt gcaggagctg ggcaagggga
     1621 gcacgcatcc gcagcccggg gtgtcgccac ccgctgcccc ggcggcgccc ggccccaagg
     1681 acggccccgg ggagacggac gcgtttggca acagcgaggg caaagagctg gtggcctcag
     1741 gtgaaaataa aataaaacag ggtctgttac ctagcttgga agatttgctg ttctatacaa
     1801 ttgctgaagg acaagagaaa atacctgttc ataaatttat tacagcactc aaatctacag
     1861 gattgcgaac gtctgatccc aggttgaaag agtgtatgga tatgttaaga ttaactcttc
     1921 aaacaacatc agatggtgtc atgctagaca aagatctttt taaaaaatgt gttcagagca
     1981 acattgtttt gttgacacaa gcatttagaa gaaagtttgt gattcctgac tttatgtctt
     2041 ttacctcaca cattgatgag ttatatgaaa gtgctaaaaa gcagtctgga ggaaaggttg
     2101 cagattatat tcctcaactg gccaaattca gtcccgattt gtggggtgtg tctgtttgta
     2161 cagtagatgg acagaggcat tctactggag ataccaaagt tcccttctgt cttcaggcct
     2221 gtgtaaaacc tttgaaatat gccattgctg ttaatgatct tggaactgaa tatgtgcatc
     2281 gatatgttgg aaaagagccg agtggactaa gattcaacaa actatttttg aatgaagatg
     2341 ataaaccaca taatcctatg gtaaatgctg gagcaattgt tgtgacttca ctaataaagc
     2401 aaggagtaaa taatgctgaa aaatttgact atgtcatgca gtttttgaat aagatggctg
     2461 gtaatgaata tgttggattc agtaatgcaa cgtttcagtc tgaaagagaa agtggagatc
     2521 gaaattttgc aataggatat tacttaaaag aaaagaagtg ttttccagaa ggcacagaca
     2581 tggttggtat attagacttc tacttccagc tgtgctccat tgaagtgact tgtgaatcag
     2641 ccagtgtgat ggctgcgaca ctggctaatg gtggtttctg cccaattact ggtgaaagag
     2701 tactgagccc tgaagcagtt cgaaatacat tgagtttgat gcattcctgt ggcatgtatg
     2761 acttctcagg gcagtttgct ttccatgttg gtcttcctgc aaaatctgga gttgctgggg
     2821 gcattctttt agttgtcccc aatgttatgg gtatgatgtg ctggtctcct cctctggata
     2881 agatgggcaa cagtgttaag ggaattcact tttgtcacga tcttgtttct ctgtgtaatt
     2941 tccataacta tgataatttg agacactttg caaaaaaact tgatcctcga agagaaggtg
     3001 gtgatcaaag ggtaaagtca gtgataaatc ttttgtttgc tgcatatact ggagatgtgt
     3061 ctgcacttcg aagatttgct ttgtcagcta tggacatgga acagcgggac tatgattcta
     3121 gaacagcact ccatgtagct gctgcagagg gtcatgttga agttgttaaa tttttgctgg
     3181 aagcctgcaa agtaaaccct ttccccaagg acaggtggaa taacactccc atggatgaag
     3241 cactgcactt tggacaccat gatgtattta aaattctcca agaataccaa gtccagtaca
     3301 cacctcaagg agattctgac aacgggaagg aaaatcaaac cgtccataag aatcttgatg
     3361 gattgttgaa gggtcaagac aattctgcag atatccagca cagtggcggc cgctcgagtc
     3421 tagagggccc gcggttcgaa ggtaagccta tccctaaccc tctcctcggt ctcgattcta
     3481 cgtgagtcga ccctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3541 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3601 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3661 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3721 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3781 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagcttacc
     3841 atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc
     3901 gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt
     3961 gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac
     4021 aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag
     4081 gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag
     4141 ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc
     4201 gaggagcagg actgaccgat cgataaaata aaagatttta tttagtctcc agaaaaaggg
     4261 gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa
     4321 ggcatggaaa aatacataac tgagaataga gaagttcaga tcaaggtcag gaacagatgg
     4381 aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg
     4441 gccaagaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc
     4501 ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtccagcc ctcagcagtt
     4561 tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt
     4621 atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc
     4681 tcaataaaag agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc
     4741 ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg
     4801 ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcacatgc
     4861 agcatgtatc aaaattaatt tggttttttt tcttaagtat ttacattaaa tggccatagt
     4921 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     4981 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5041 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5101 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5161 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5221 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5281 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5341 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5401 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5461 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5521 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5581 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5641 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5701 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     5761 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5821 gattatcaaa aaggatcttc acctagatcc ttttgcggcc gcaaatcaat ctaaagtata
     5881 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5941 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6001 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6061 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6121 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6181 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6241 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6301 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6361 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6421 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6481 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6541 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6601 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6661 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6721 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6781 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6841 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgcagc
//
