LOCUS       sequence-205854-        6037 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6037)
  AUTHORS   Scott KL, Kabbarah O, Liang MC, Ivanova E, Anagnostou V, Wu J, 
            Dhakal S, Wu M, Chen S, Feinberg T, Huang J, Saci A, Widlund HR, 
            Fisher DE, Xiao Y, Rimm DL, Protopopov A, Wong KK, Chin L
  TITLE     GOLPH3 modulates mTOR signalling and rapamycin sensitivity in 
            cancer.
  JOURNAL   Nature. 2009 Jun 25. 459(7250):1085-90.
  PUBMED    19553991
REFERENCE   2  (bases 1 to 6037)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2009 Jun 25. 459(7250):1085-90."
COMMENT     Sequence Label: pBABE-HA-GOLPH3
FEATURES             Location/Qualifiers
     source          1..6037
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             39..455
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     385..407
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     423..439
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             475..501
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             667..681
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     primer_bind     complement(1464..1484)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1469..1798
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1649..1784
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1808..2407
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1808..1827)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2304..2324
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2628..3097
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3227..3244)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3398..3986)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3478..3497)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4146..4937,4938..5006))
                     /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     4769..4788
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5007..5111)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5142..5611
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    5676..5875
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 tctgtctgac tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct
       61 taagtttgac cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag
      121 atgtcaagaa gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg
      181 gatggccgcg agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct
      241 tttcacctgg cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct
      301 tggcttttga cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc
      361 ttcctccatc cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct
      421 ccctttatcc agccctcact ccttctctag gcgccggccg gatctgccac catgtatcca
      481 tatgatgttc cagattatgc tggatccccc gggctgcagg aattccggac ctcgctgacc
      541 cagcgcagct ccggcctggt gcagcggcgc accgaggcct cccgcaacgc cgccgacaag
      601 gagcgggcgg cgggcggcgg cgccggcagc agcgaggacg acgcgcagag ccgccgcgac
      661 gagcaggacg acgacgacaa gggcgactcc aaggaaacgc ggctgaccct gatggaggaa
      721 gtgctcctgc tgggcctcaa ggaccgcgag ggttacacat cattttggaa tgactgtata
      781 tcatctggat tacgtggctg tatgttaatt gaattagcat tgagaggaag gttacaacta
      841 gaggcttgtg gaatgagacg taaaagtcta ttaacaagaa aggtaatctg taagtcagat
      901 gctccaacag gggatgttct tcttgatgaa gctctgaagc atgttaagga aactcagcct
      961 ccagaaacgg tccagaactg gattgaatta cttagtggtg agacatggaa tccattaaaa
     1021 ttgcattatc agttaagaaa tgtacgggaa cgattagcta aaaacctggt ggaaaagggt
     1081 gtattgacaa cagagaaaca gaacttccta ctttttgaca tgacaacaca tcccctcacc
     1141 aataacaaca ttaagcagcg cctcatcaag aaagtacagg aagccgttct tgacaaatgg
     1201 gtgaatgacc ctcaccgcat ggacaggcgc ttgctggccc tcatttacct ggctcatgcc
     1261 tcggacgtcc tggagaatgc ttttgctcct cttctggacg agcagtatga tttggctacc
     1321 aagagagtgc ggcagcttct cgacttagac cctgaagtgg aatgtctgaa ggccaacacc
     1381 aatgaggttc tgtgggcggt ggtggcggcg ttcaccaagt aaccggaatt cgatatcaag
     1441 cttatcgata ccgtcgaccc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     1501 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     1561 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     1621 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     1681 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     1741 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     1801 gcttaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag
     1861 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     1921 tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     1981 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     2041 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     2101 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     2161 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg
     2221 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     2281 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     2341 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     2401 tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa aggagcgcac gaccccatgc
     2461 atcgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac
     2521 ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata
     2581 actgagaata gagaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc
     2641 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac
     2701 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     2761 aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat
     2821 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     2881 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
     2941 acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc
     3001 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
     3061 tctgagtgat tgactacccg tcagcggggg tctttcacat gcagcatgta tcaaaattaa
     3121 tttggttttt tttcttaagt atttacatta aatggccata gttgcattaa tgaatcggcc
     3181 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     3241 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3301 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3361 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3421 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3481 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3541 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3601 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3661 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3721 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3781 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3841 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3901 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3961 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4021 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4081 tcacctagat ccttttgcgg ccgcaaatca atctaaagta tatatgagta aacttggtct
     4141 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4201 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4261 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4321 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4381 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4441 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4501 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4561 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4621 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4681 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4741 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4801 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4861 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4921 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4981 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5041 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5101 ggggttccgc gcacatttcc ccgaaaagtg ccacctgcag cctgaatatg ggccaaacag
     5161 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga
     5221 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac
     5281 agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc
     5341 agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc
     5401 ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct
     5461 cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa
     5521 ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag
     5581 tgattgacta cccgtcagcg ggggtctttc atttgggggc tcgtccggga tcgggagacc
     5641 cctgcccagg gaccaccgac ccaccaccgg gaggcaagct ggccagcaac ttatctgtgt
     5701 ctgtccgatt gtctagtgtc tatgactgat tttatgcgcc tgcgtcggta ctagttagct
     5761 aactagctct gtatctggcg gacccgtggt ggaactgacg agttctgaac acccggccgc
     5821 aaccctggga gacgtcccag ggactttggg ggccgttttt gtggcccgac ctgaggaagg
     5881 gagtcgatgt ggaatccgac cccgtcagga tatgtggttc tggtaggaga cgagaaccta
     5941 aaacagttcc cgcctccgtc tgaatttttg ctttcggttt ggaaccgaag ccgcgcgtct
     6001 tgtctgctgc agcgctgcag catcgttctg tgttgtc
//
