LOCUS       62056_12780        8917 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8917)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8917
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1442..1645
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             1751..3400
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     promoter        3444..3655
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3668..3936
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3992..4657
                     /codon_start=1
                     /label=CopGFP
                     /note="green fluorescent protein 2 from Pontellina plumata,
                     also known as ppluGFP2 (Shagin et al., 2004)"
                     /translation="LPAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGAL
                     TFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYR
                     YEAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDG
                     GYYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA
                     "
     CDS             4727..4780
                     /codon_start=1
                     /label=T2A
                     /note="2A peptide from Thosea asigna virus capsid protein"
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4781..5377
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    5488..5620
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6896..6912)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6920..6936)
                     /label=lac operator
                     /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(6944..6974)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6989..7010)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7298..7886)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8060..8917)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgc gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaaa caaaaactca
     1081 aaatttcttc tataaagtaa caaaactttt atgagggaca gccccccccc aaagccccca
     1141 gggatgtaat tacgtccctc ccccgctagg gggcagcagc gagccgcccg gggctccgct
     1201 ccggtccggc gctccccccg catccccgag ccggcagcgt gcggggacag cccgggcacg
     1261 gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag
     1321 acacctgggg ggatacgggg aaaaggcctc caaggcctac tagtattatg cccagtacat
     1381 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     1441 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     1501 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     1561 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     1621 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca
     1681 tccacgctgt tttgacctcc atagaagatt ctagagctag cgaattcgaa tttaaatcgg
     1741 atccgccacc atggaagacg ccaaaaacat aaagaaaggc ccggcgccat tctatccgct
     1801 ggaagatgga accgctggag agcaactgca taaggctatg aagagatacg ccctggttcc
     1861 tggaacaatt gcttttacag atgcacatat cgaggtggac atcacttacg ctgagtactt
     1921 cgaaatgtcc gttcggttgg cagaagctat gaaacgatat gggctgaata caaatcacag
     1981 aatcgtcgta tgcagtgaaa actctcttca attctttatg ccggtgttgg gcgcgttatt
     2041 tatcggagtt gcagttgcgc ccgcgaacga catttataat gaacgtgaat tgctcaacag
     2101 tatgggcatt tcgcagccta ccgtggtgtt cgtttccaaa aaggggttgc aaaaaatttt
     2161 gaacgtgcaa aaaaagctcc caatcatcca aaaaattatt atcatggatt ctaaaacgga
     2221 ttaccaggga tttcagtcga tgtacacgtt cgtcacatct catctacctc ccggttttaa
     2281 tgaatacgat tttgtgccag agtccttcga tagggacaag acaattgcac tgatcatgaa
     2341 ctcctctgga tctactggtc tgcctaaagg tgtcgctctg cctcatagaa ctgcctgcgt
     2401 gagattctcg catgccagag atcctatttt tggcaatcaa atcattccgg atactgcgat
     2461 tttaagtgtt gttccattcc atcacggttt tggaatgttt actacactcg gatatttgat
     2521 atgtggattt cgagtcgtct taatgtatag atttgaagaa gagctgtttc tgaggagcct
     2581 tcaggattac aagattcaaa gtgcgctgct ggtgccaacc ctattctcct tcttcgccaa
     2641 aagcactctg attgacaaat acgatttatc taatttacac gaaattgctt ctggtggcgc
     2701 tcccctctct aaggaagtcg gggaagcggt tgccaagagg ttccatctgc caggtatcag
     2761 gcaaggatat gggctcactg agactacatc agctattctg attacacccg agggggatga
     2821 taaaccgggc gcggtcggta aagttgttcc attttttgaa gcgaaggttg tggatctgga
     2881 taccgggaaa acgctgggcg ttaatcaaag aggcgaactg tgtgtgagag gtcctatgat
     2941 tatgtccggt tatgtaaaca atccggaagc gaccaacgcc ttgattgaca aggatggatg
     3001 gctacattct ggagacatag cttactggga cgaagacgaa cacttcttca tcgttgaccg
     3061 cctgaagtct ctgattaagt acaaaggcta tcaggtggct cccgctgaat tggaatccat
     3121 cttgctccaa caccccaaca tcttcgacgc aggtgtcgca ggtcttcccg acgatgacgc
     3181 cggtgaactt cccgccgccg ttgttgtttt ggagcacgga aagacgatga cggaaaaaga
     3241 gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt
     3301 gtttgtggac gaagtaccga aaggtcttac cggaaaactc gacgcaagaa aaatcagaga
     3361 gatcctcata aaggccaaga agggcggaaa gatcgccgtg taagcggccg caaggatctg
     3421 cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
     3481 ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga
     3541 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
     3601 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa
     3661 gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc
     3721 acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc
     3781 cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag
     3841 cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt
     3901 ctttgtttcg ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct
     3961 agacgccacc atggagagcg acgagagcgg cctgcccgcc atggagatcg agtgccgcat
     4021 caccggcacc ctgaacggcg tggagttcga gctggtgggc ggcggagagg gcacccccaa
     4081 gcagggccgc atgaccaaca agatgaagag caccaaaggc gccctgacct tcagccccta
     4141 cctgctgagc cacgtgatgg gctacggctt ctaccacttc ggcacctacc ccagcggcta
     4201 cgagaacccc ttcctgcacg ccatcaacaa cggcggctac accaacaccc gcatcgagaa
     4261 gtacgaggac ggcggcgtgc tgcacgtgag cttcagctac cgctacgagg ccggccgcgt
     4321 gatcggcgac ttcaaggtgg tgggcaccgg cttccccgag gacagcgtga tcttcaccga
     4381 caagatcatc cgcagcaacg ccaccgtgga gcacctgcac cccatgggcg ataacgtgct
     4441 ggtgggcagc ttcgcccgca ccttcagcct gcgcgacggc ggctactaca gcttcgtggt
     4501 ggacagccac atgcacttca agagcgccat ccaccccagc atcctgcaga acgggggccc
     4561 catgttcgcc ttccgccgcg tggaggagct gcacagcaac accgagctgg gcatcgtgga
     4621 gtaccagcac gccttcaaga cccccatcgc cttcgccaga tcccgcgctc agtcgtccaa
     4681 ttctgccgtg gacggcaccg ccggacccgg ctccaccgga tctcgcgagg gcagaggaag
     4741 tcttctaaca tgcggtgacg tggaggagaa tcccggccct atgaccgagt acaagcccac
     4801 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     4861 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     4921 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     4981 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     5041 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     5101 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     5161 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     5221 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     5281 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     5341 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga aatcaacctc tggattacaa
     5401 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     5461 cgctgcttta atgcctttgt atcagttaac ttgtttattg cagcttataa tggttacaaa
     5521 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5581 ggtttgtcca aactcatcaa tgtatcttat catgtctgga attgactcaa atgatgtcaa
     5641 ttagtctatc agaagctatc tggtctccct tccgggggac aagacatccc tgtttaatat
     5701 ttaaacagca gtgttcccaa actgggttct tatatccctt gctctggtca accaggttgc
     5761 agggtttcct gtcctcacag gaacgaagtc cctaaagaaa cagtggcagc caggtttagc
     5821 cccggaattg actggattcc ttttttaggg cccattggta tggctttttc cccgtatccc
     5881 cccaggtgtc tgcaggctca aagagcagcg agaagcgttc agaggaaagc gatcccgtgc
     5941 caccttcccc gtgcccgggc tgtccccgca cgctgccggc tcggggatgc ggggggagcg
     6001 ccggaccgga gcggagcccc gggcggctcg ctgctgcccc ctagcggggg agggacgtaa
     6061 ttacatccct gggggctttg ggggggggct gtccctgata tctataacaa gaaaatatat
     6121 atataataag ttatcacgta agtagaacat gaaataacaa tataattatc gtatgagtta
     6181 aatcttaaaa gtcacgtaaa agataatcat gcgtcatttt gactcacgcg gtcgttatag
     6241 ttcaaaatca gtgacactta ccgcattgac aagcacgcct cacgggagct ccaagcggcg
     6301 actgagatgt cctaaatgca cagcgacgga ttcgcgctat ttagaaagag agagcaatat
     6361 ttcaagaatg catgcgtcaa ttttacgcag actatctttc tagggttaat ctagctgcat
     6421 caggatcata tcgtcgggtc ttttttccgg ctcagtcatc gcccaagctg gcgctatctg
     6481 ggcatcgggg aggaagaagc ccgtgccttt tcccgcgagg ttgaagcggc atggaaagag
     6541 tttgccgagg atgactgctg ctgcattgac gttgagcgaa aacgcacgtt taccatgatg
     6601 attcgggaag gtgtggccat gcacgccttt aacggtgaac tgttcgttca ggccacctgg
     6661 gataccagtt cgtcgcggct tttccggaca cagttccgga tggtcagccc gaagcgcatc
     6721 agcaacccga acaataccgg cgacagccgg aactgccgtg ccggtgtgca gattaatgac
     6781 agcggtgcgg cgctgggata ttacgtcagc gaggacgggt atcctggctg gatgccgcag
     6841 aaatggacat ggataccccg tgagttaccc ggcgggcgcg cttggcgtaa tcatggtcat
     6901 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     6961 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     7021 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     7081 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     7141 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     7201 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     7261 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     7321 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     7381 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     7441 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     7501 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     7561 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     7621 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7681 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     7741 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7801 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7861 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7921 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7981 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     8041 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     8101 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     8161 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     8221 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     8281 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     8341 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     8401 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     8461 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     8521 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     8581 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     8641 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     8701 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     8761 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     8821 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     8881 agggaataag ggcgacacgg aaatgttgaa tactcat
//
