LOCUS       sequence-174189-        6634 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 6634)
  TITLE     Akt
REFERENCE   2  (bases 1 to 6634)
  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"
COMMENT     Sequence Label: 1271 pBabe puroL Myr HA Akt2
FEATURES             Location/Qualifiers
     source          1..6634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             17..486
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    551..750
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             951..1367
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1297..1319
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1335..1351
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1399..1440
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSKPKDPSQR"
     CDS             1444..1470
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YAYDVPDYA"
     CDS             1450..1476
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     primer_bind     complement(2973..2993)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2978..3307
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3158..3293
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3317..3916
                     /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(3317..3336)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3813..3833
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4137..4606
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4736..4753)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4907..5495)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4987..5006)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5655..6446,6447..6515))
                     /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     6278..6297
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6516..6620)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aagtgccacc tgcagcctga atatgggcca aacaggatat ctgtggtaag cagttcctgc
       61 cccggctcag ggccaagaac agatggaaca gctgaatatg ggccaaacag gatatctgtg
      121 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtccagc
      181 cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga
      241 ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct
      301 gctccccgag ctcaataaaa gagcccacaa cccctcactc ggggcgccag tcctccgatt
      361 gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg
      421 tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt
      481 ctttcatttg ggggctcgtc cgggatcggg agacccctgc ccagggacca ccgacccacc
      541 accgggaggc aagctggcca gcaacttatc tgtgtctgtc cgattgtcta gtgtctatga
      601 ctgattttat gcgcctgcgt cggtactagt tagctaacta gctctgtatc tggcggaccc
      661 gtggtggaac tgacgagttc tgaacacccg gccgcaaccc tgggagacgt cccagggact
      721 ttgggggccg tttttgtggc ccgacctgag gaagggagtc gatgtggaat ccgaccccgt
      781 caggatatgt ggttctggta ggagacgaga acctaaaaca gttcccgcct ccgtctgaat
      841 ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct gctgcagcgc tgcagcatcg
      901 ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc tgaaaattag ggccagactg
      961 ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg atcgctcaca
     1021 accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca gaatggccaa
     1081 cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc acccaggtta
     1141 agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc tacatcgtga
     1201 cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta caccctaagc
     1261 ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct cgttcgaccc
     1321 cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccggc cggatcgaag
     1381 actaagcttc ccaccaccat ggggagcagc aagagcaagc ccaaggaccc cagccagcgc
     1441 gcctacgcct acgacgtgcc cgactacgcc tccctcggat ccatgaatga ggtgtctgtc
     1501 atcaaagaag gctggctcca caagcgtggt gaatacatca agacctggag gccacggtac
     1561 ttcctgctga agagcgacgg ctccttcatt gggtacaagg agaggcccga ggcccctgat
     1621 cagactctac cccccttaaa caacttctcc gtagcagaat gccagctgat gaagaccgag
     1681 aggccgcgac ccaacacctt tgtcatacgc tgcctgcagt ggaccacagt catcgagagg
     1741 accttccacg tggattctcc agacgagagg gaggagtgga tgcgggccat ccagatggtc
     1801 gccaacagcc tcaagcagcg ggccccaggc gaggacccca tggactacaa gtgtggctcc
     1861 cccagtgact cctccacgac tgaggagatg gaagtggcgg tcagcaaggc acgggctaaa
     1921 gtgaccatga atgacttcga ctatctcaaa ctccttggca agggaacctt tggcaaagtc
     1981 atcctggtgc gggagaaggc cactggccgc tactacgcca tgaagatcct gcggaaggaa
     2041 gtcatcattg ccaaggatga agtcgctcac acagtcaccg agagccgggt cctccagaac
     2101 accaggcacc cgttcctcac tgcgctgaag tatgccttcc agacccacga ccgcctgtgc
     2161 tttgtgatgg agtatgccaa cgggggtgag ctgttcttcc acctgtcccg ggagcgtgtc
     2221 ttcacagagg agcgggcccg gttttatggt gcagagattg tctcggctct tgagtacttg
     2281 cactcgcggg acgtggtata ccgcgacatc aagctggaaa acctcatgct ggacaaagat
     2341 ggccacatca agatcactga ctttggcctc tgcaaagagg gcatcagtga cggggccacc
     2401 atgaaaacct tctgtgggac cccggagtac ctggcgcctg aggtgctgga ggacaatgac
     2461 tatggccggg ccgtggactg gtgggggctg ggtgtggtca tgtacgagat gatgtgcggc
     2521 cgcctgccct tctacaacca ggaccacgag cgcctcttcg agctcatcct catggaagag
     2581 atccgcttcc cgcgcacgct cagccccgag gccaagtccc tgcttgctgg gctgcttaag
     2641 aaggacccca agcagaggct tggtgggggg cccagcgatg ccaaggaggt catggagcac
     2701 aggttcttcc tcagcatcaa ctggcaggac gtggtccaga agaagctcct gccacccttc
     2761 aaacctcagg tcacgtccga ggtcgacaca aggtacttcg atgatgaatt taccgcccag
     2821 tccatcacaa tcacaccccc tgaccgctat gacagcctgg gcttactgga gctggaccag
     2881 cggacccact tcccccagtt ctcctactcg gccagcatcc gcgagtgaat tcgatatctc
     2941 gagtaattcg ccagcacagt ggtcgaccct gtggaatgtg tgtcagttag ggtgtggaaa
     3001 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3061 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3121 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3181 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3241 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3301 ttgcaaaaag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     3361 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     3421 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     3481 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     3541 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     3601 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     3661 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     3721 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     3781 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     3841 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     3901 caagcccggt gcctgacgcc cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg
     3961 accccatgca tcgataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa
     4021 agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa
     4081 aaatacataa ctgagaatag agaagttcag atcaaggtca ggaacagatg gaacagctga
     4141 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac
     4201 agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
     4261 ctcagggcca agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagagaa
     4321 ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact
     4381 aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa
     4441 gagcccacaa cccctcactc ggggcgccag tcctccgatt gactgagtcg cccgggtacc
     4501 cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg
     4561 agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcacatg cagcatgtat
     4621 caaaattaat ttggtttttt ttcttaagta tttacattaa atggccatag ttgcattaat
     4681 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     4741 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4801 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4861 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4921 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4981 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5041 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5101 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5161 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5221 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5281 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5341 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5401 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5461 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5521 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5581 aaaggatctt cacctagatc cttttgcggc cgcaaatcaa tctaaagtat atatgagtaa
     5641 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5701 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5761 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5821 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5881 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5941 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6001 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6061 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6121 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6181 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6241 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6301 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6361 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6421 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6481 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6541 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6601 aaacaaatag gggttccgcg cacatttccc cgaa
//
