LOCUS       Exported                8611 bp ds-DNA     circular SYN 06-MAR-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8611)
  AUTHORS   MReijns
  TITLE     Direct Submission
  JOURNAL   Exported Mar 6, 2018 from SnapGene Viewer 4.1.7
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8611
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    580..921
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             988..1404
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     CDS             1429..2145
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     protein_bind    2218..2242
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2245..3768
                     /codon_start=1
                     /label=mcGAS
                     /translation="MEDPRRRTTAPRAKKPSAKRAPTQPSRTRAHAESCGPQRGARSRR
                     AERDGDTTEKPRAPGPRVHPARATELTKDAQPSAMDAAGATARPAVRVPQQQAILDPEL
                     PAVREPQPPADPEARKVVRGPSHRRGARSTGQPRAPRGSRKEPDKLKKVLDKLRLKRKD
                     ISEAAETVNKVVERLLRRMQKRESEFKGVEQLNTGSYYEHVKISAPNEFDVMFKLEVPR
                     IELQEYYETGAFYLVKFKRIPRGNPLSHFLEGEVLSATKMLSKFRKIIKEEVKEIKDID
                     VSVEKEKPGSPAVTLLIRNPEEISVDIILALESKGSWPISTKEGLPIQGWLGTKVRTNL
                     RREPFYLVPKNAKDGNSFQGETWRLSFSHTEKYILNNHGIEKTCCESSGAKCCRKECLK
                     LMKYLLEQLKKEFQELDAFCSYHVKTAIFHMWTQDPQDSQWDPRNLSSCFDKLLAFFLE
                     CLRTEKLDHYFIPKFNLFSQELIDRKSKEFLSKKIEYERNNGFPIFDKL"
     promoter        3785..4284
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             4305..4904
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     LTR             5004..5518
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5680..5696
                     /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(5704..5734)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5749..5770
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6058..6646)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6817..7677)
                     /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"
     promoter        complement(7678..7782)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca
       61 tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc
      121 agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
      181 aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt
      241 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
      301 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
      361 cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tattcccaat
      421 aaagcctctt gctgtttgca tccgaatcgt ggactcgctg atccttggga gggtctcctc
      481 agattgattg actgcccacc tcgggggtct ttcatttgga ggttccaccg agatttggag
      541 acccctgccc agggaccacc gacccccccg ccgggaggta agctggccag cggtcgtttc
      601 gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg
      661 tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttctga
      721 acacccggcc gcaaccctgg gagacgtccc agggactttg ggggccgttt ttgtggcccg
      781 acctgaggaa gggagtcgat gtggaatccg accccgtcag gatatgtggt tctggtagga
      841 gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttggaaccga
      901 agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact
      961 gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc
     1021 ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag
     1081 agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga
     1141 gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc
     1201 ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac
     1261 ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc
     1321 gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca
     1381 gccctcactc cttctctagg cgccggaatt agatctctcg aggttaacat ggtgagcaag
     1441 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     1501 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     1561 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     1621 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     1681 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     1741 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1801 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1861 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     1921 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     1981 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     2041 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     2101 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtccgg ccggactcag
     2161 atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg gatcatcaca
     2221 agtttgtaca aaaaagcagg cttcatggaa gatccgcgta gaaggacgac ggcgccacgc
     2281 gctaagaagc cgtccgcgaa gcgcgccccg acgcagccca gcaggaccag ggcccacgcg
     2341 gaaagctgcg gcccgcaaag gggggctcga tcgcggcggg cggagcgtga cggggacacc
     2401 acggagaagc cacgtgcccc agggccccga gtgcatccag caagggccac tgagctcacc
     2461 aaagatgcac agccctcggc catggacgcg gcaggagcca ccgcgcggcc tgccgtccgg
     2521 gtgccccagc agcaggccat cctggatccg gagctgcccg ccgtacggga gccccagccg
     2581 cccgcggatc ccgaggcgcg gaaagtcgta aggggaccta gccacagaag gggcgcgcgc
     2641 tccaccgggc agcccagagc gccgcgaggg tccaggaagg aaccggacaa gctaaagaag
     2701 gtgctggaca aattgagatt gaaacgcaaa gatatctcgg aggcggccga gacggtgaat
     2761 aaagttgtgg aacgcctgct gcgcagaatg cagaaacggg agtcggagtt caaaggtgtg
     2821 gagcagctga acactggcag ctactatgaa catgtgaaga tttctgctcc taatgaattt
     2881 gatgttatgt ttaaactgga agtccccagg attgagctac aagaatatta tgaaacaggt
     2941 gctttctatc ttgtgaaatt caaaagaatt ccacgaggaa atccgctgag tcatttctta
     3001 gaaggggaag tattatcagc taccaagatg ctgtcaaagt ttaggaaaat cattaaagaa
     3061 gaagttaaag aaatcaaaga tatagatgtc agtgtggaga aggaaaaacc aggaagccct
     3121 gctgtaacac ttcttatcag gaaccctgaa gaaatctctg tggatataat tctggctttg
     3181 gagtcaaaag gcagctggcc tattagtacc aaagaaggac tacctattca aggctggctg
     3241 ggcacaaaag tgaggaccaa tctaagacga gagccgtttt atctcgtacc caagaatgca
     3301 aaggatggaa atagttttca aggagagacc tggcgcctct ctttctctca cactgaaaag
     3361 tacattttga ataatcacgg gatagagaaa acatgctgtg aatcttccgg agcaaaatgc
     3421 tgcagaaaag aatgtttaaa attaatgaaa taccttttgg aacagttgaa aaaagagttt
     3481 caagagctgg atgcattctg ttcctaccat gtgaaaactg ccatctttca catgtggacc
     3541 caggacccgc aggacagtca gtgggacccc aggaacctca gctcctgctt cgataagttg
     3601 ttagcattct ttcttgagtg cctcaggaca gagaaactgg atcattattt tattccaaag
     3661 ttcaatctat tctctcaaga actaattgac cgaaaaagta aagaatttct atcgaagaaa
     3721 attgaatatg aaagaaataa tgggtttcca atttttgaca agctttgagt taacgaattc
     3781 taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg
     3841 ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt
     3901 aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta
     3961 gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta
     4021 gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc
     4081 ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga
     4141 aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg
     4201 tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc
     4261 tcttcctcat ctccgggcct ttcgacctgc agcccaagct taccatgacc gagtacaagc
     4321 ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg
     4381 cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg
     4441 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg
     4501 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
     4561 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
     4621 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
     4681 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc
     4741 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
     4801 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
     4861 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacgcccg ccccacgacc
     4921 cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc gataaaataa aagattttat
     4981 ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt
     5041 aagtaacgcc attttgcaag gcatggaaaa tacataactg agaatagaga agttcagatc
     5101 aaggttagga acagagagac agcagaatat gggccaaaca ggatatctgt ggtaagcagt
     5161 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtcccg ccctcagcag
     5221 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
     5281 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
     5341 gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat agactgcgtc
     5401 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
     5461 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcatgg
     5521 gtaacagttt cttgaagttg gagaacaaca ttctgagggt aggagtcgaa tattaagtaa
     5581 tcctgactca attagccact gttttgaatc cacatactcc aatactcctg aaatagttca
     5641 ttatggacag cgcagaaaga gctggggaga attgtgaaat tgttatccgc tcacaattcc
     5701 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5761 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5821 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5881 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5941 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6001 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6061 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6121 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6181 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6241 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6301 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6361 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6421 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6481 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6541 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6601 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6661 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6721 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6781 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6841 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6901 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6961 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7021 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7081 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7141 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7201 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7261 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7321 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7381 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7441 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7501 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7561 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7621 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7681 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7741 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7801 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     7861 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     7921 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     7981 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     8041 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     8101 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     8161 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     8221 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcgc aaggaatggt
     8281 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
     8341 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
     8401 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
     8461 gcgtagaggc gattagtcca atttgttaaa gacaggatat cagtggtcca ggctctagtt
     8521 ttgactcaac aatatcacca gctgaagcct atagagtacg agccatagat aaaataaaag
     8581 attttattta gtctccagaa aaagggggga a
//
