LOCUS       Exported                7219 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    MSCV H-Ras V12 IRES GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7219)
  TITLE     Lowe Lab Plasmids
REFERENCE   2  (bases 1 to 7219)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7219
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(455..1315)
                     /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     1078..1097
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1316..1420)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1488..1506
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1544..1566)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1666..1685
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1879..1901
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2029..2048
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             2248..2764
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    2828..3169
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             3236..3652
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3582..3604
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3620..3636
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             3709..4278
                     /codon_start=1
                     /product="human oncoprotein generated by the G12V mutation 
                     in the small GTPase H-Ras"
                     /label=H-Ras (G12V)
                     /translation="MTEYKLVVVGAVGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHQYREQIKRVK
                     DSDDVPMVLVGNKCDLAARTVESRQAQDLARSYGIPYIETSAKTRQGVEDAFYTLVREI
                     RQHKLRKLNPPDESGPGCMSCKCVLS"
     primer_bind     complement(4572..4589)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4799..4818
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4980..5699
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             4980..5696
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5025..5046)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5286..5305)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5633..5654
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             5861..6375
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    6537..6553
                     /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(6561..6591)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6606..6627
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6744..6761)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6915..284)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6995..7014)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
       61 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
      121 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
      181 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
      241 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
      301 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
      361 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
      421 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
      481 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
      541 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
      601 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
      661 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
      721 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
      781 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
      841 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
      901 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
      961 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     1021 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     1081 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     1141 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     1201 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     1261 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     1321 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     1381 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     1441 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     1501 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     1561 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     1621 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     1681 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     1741 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     1801 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     1861 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggcgcaa ggaatggtgc
     1921 atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
     1981 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
     2041 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
     2101 gtagaggcga ttagtccaat ttgttaaaga caggatatca gtggtccagg ctctagtttt
     2161 gactcaacaa tatcaccagc tgaagcctat agagtacgag ccatagataa aataaaagat
     2221 tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct
     2281 agcttaagta acgccatttt gcaaggcatg gaaaatacat aactgagaat agagaagttc
     2341 agatcaaggt taggaacaga gagacagcag aatatgggcc aaacaggata tctgtggtaa
     2401 gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg tcccgccctc
     2461 agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct
     2521 gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc
     2581 cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgatagact
     2641 gcgtcgcccg ggtacccgta ttcccaataa agcctcttgc tgtttgcatc cgaatcgtgg
     2701 actcgctgat ccttgggagg gtctcctcag attgattgac tgcccacctc gggggtcttt
     2761 catttggagg ttccaccgag atttggagac ccctgcccag ggaccaccga cccccccgcc
     2821 gggaggtaag ctggccagcg gtcgtttcgt gtctgtctct gtctttgtgc gtgtttgtgc
     2881 cggcatctaa tgtttgcgcc tgcgtctgta ctagttagct aactagctct gtatctggcg
     2941 gacccgtggt ggaactgacg agttctgaac acccggccgc aaccctggga gacgtcccag
     3001 ggactttggg ggccgttttt gtggcccgac ctgaggaagg gagtcgatgt ggaatccgac
     3061 cccgtcagga tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc
     3121 tgaatttttg ctttcggttt ggaaccgaag ccgcgcgtct tgtctgctgc agcgctgcag
     3181 catcgttctg tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa attagggcca
     3241 gactgttacc actcccttaa gtttgacctt aggtcactgg aaagatgtcg agcggatcgc
     3301 tcacaaccag tcggtagatg tcaagaagag acgttgggtt accttctgct ctgcagaatg
     3361 gccaaccttt aacgtcggat ggccgcgaga cggcaccttt aaccgagacc tcatcaccca
     3421 ggttaagatc aaggtctttt cacctggccc gcatggacac ccagaccagg tcccctacat
     3481 cgtgacctgg gaagccttgg cttttgaccc ccctccctgg gtcaagccct ttgtacaccc
     3541 taagcctccg cctcctcttc ctccatccgc cccgtctctc ccccttgaac ctcctcgttc
     3601 gaccccgcct cgatcctccc tttatccagc cctcactcct tctctaggcg ccggaattag
     3661 atcccagtgt ggtggtacag cttggggggc cagggccggg gccgagcgat gacggaatat
     3721 aagctggtgg tggtgggcgc cgtcggtgtg ggcaagagtg cgctgaccat ccagctgatc
     3781 cagaaccatt ttgtggacga atacgacccc actatagagg attcctaccg gaagcaggtg
     3841 gtcattgatg gggagacgtg cctgttggac atcctggata ccgccggcca ggaggagtac
     3901 agcgccatgc gggaccagta catgcgcacc ggggagggct tcctgtgtgt gtttgccatc
     3961 aacaacacca agtcttttga ggacatccac cagtacaggg agcagatcaa acgggtgaag
     4021 gactcggatg acgtgcccat ggtgctggtg gggaacaagt gtgacctggc tgcacgcact
     4081 gtggaatctc ggcaggctca ggacctcgcc cgaagctacg gcatccccta catcgagacc
     4141 tcggccaaga cccggcaggg agtggaggat gccttctaca cgttggtgcg tgagatccgg
     4201 cagcacaagc tgcggaagct gaaccctcct gatgagagtg gccccggctg catgagctgc
     4261 aagtgtgtgc tctcctgacg cagcacaagc tcaggacatg gaggtgccgg atgcaggaag
     4321 gaggtgcaga cggaaggagg aggaaggaag gacggaagca aggaaggaag gaagggctgc
     4381 tggagcccag tcacccccca agctgtagga attccgcccc ccccctaacg ttactggccg
     4441 aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc
     4501 gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag
     4561 gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt
     4621 tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa
     4681 ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc
     4741 aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg
     4801 gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat
     4861 gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa
     4921 acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac gataatacca
     4981 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     5041 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     5101 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     5161 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     5221 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     5281 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     5341 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     5401 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     5461 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     5521 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     5581 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     5641 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaaa
     5701 gcggccgcga ctctagagtc gacctgcagg catgcaagct tcaggtagcc ggctaacgtt
     5761 aacaaccggt acctctagaa ctatagctag catgcgcaaa tttaaagcgc tgatatcgat
     5821 aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag
     5881 gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaatac ataactgaga
     5941 atagagaagt tcagatcaag gttaggaaca gagagacagc agaatatggg ccaaacagga
     6001 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc
     6061 ggtcccgccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc
     6121 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
     6181 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc
     6241 ctccgataga ctgcgtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat
     6301 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca
     6361 gcgggggtct ttcatgggta acagtttctt gaagttggag aacaacattc tgagggtagg
     6421 agtcgaatat taagtaatcc tgactcaatt agccactgtt ttgaatccac atactccaat
     6481 actcctgaaa tagttcatta tggacagcgc agaaagagct ggggagaatt gtgaaattgt
     6541 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     6601 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6661 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     6721 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     6781 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     6841 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6901 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6961 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     7021 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     7081 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     7141 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     7201 gccttatccg gtaactatc
//
