LOCUS       62056_12785        7591 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 7591)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7591
                     /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"
     CDS             3401..3454
                     /codon_start=1
                     /label=T2A
                     /note="2A peptide from Thosea asigna virus capsid protein"
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3455..4051
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    4162..4294
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5570..5586)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5594..5610)
                     /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(5618..5648)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5663..5684)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5972..6560)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6734..7591)
                     /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 gagggcagag gaagtcttct
     3421 aacatgcggt gacgtggagg agaatcccgg ccctatgacc gagtacaagc ccacggtgcg
     3481 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     3541 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     3601 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     3661 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     3721 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     3781 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     3841 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     3901 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     3961 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     4021 cacctggtgc atgacccgca agcccggtgc ctgaaatcaa cctctggatt acaaaatttg
     4081 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     4141 tttaatgcct ttgtatcagt taacttgttt attgcagctt ataatggtta caaataaagc
     4201 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4261 tccaaactca tcaatgtatc ttatcatgtc tggaattgac tcaaatgatg tcaattagtc
     4321 tatcagaagc tatctggtct cccttccggg ggacaagaca tccctgttta atatttaaac
     4381 agcagtgttc ccaaactggg ttcttatatc ccttgctctg gtcaaccagg ttgcagggtt
     4441 tcctgtcctc acaggaacga agtccctaaa gaaacagtgg cagccaggtt tagccccgga
     4501 attgactgga ttcctttttt agggcccatt ggtatggctt tttccccgta tccccccagg
     4561 tgtctgcagg ctcaaagagc agcgagaagc gttcagagga aagcgatccc gtgccacctt
     4621 ccccgtgccc gggctgtccc cgcacgctgc cggctcgggg atgcgggggg agcgccggac
     4681 cggagcggag ccccgggcgg ctcgctgctg ccccctagcg ggggagggac gtaattacat
     4741 ccctgggggc tttggggggg ggctgtccct gatatctata acaagaaaat atatatataa
     4801 taagttatca cgtaagtaga acatgaaata acaatataat tatcgtatga gttaaatctt
     4861 aaaagtcacg taaaagataa tcatgcgtca ttttgactca cgcggtcgtt atagttcaaa
     4921 atcagtgaca cttaccgcat tgacaagcac gcctcacggg agctccaagc ggcgactgag
     4981 atgtcctaaa tgcacagcga cggattcgcg ctatttagaa agagagagca atatttcaag
     5041 aatgcatgcg tcaattttac gcagactatc tttctagggt taatctagct gcatcaggat
     5101 catatcgtcg ggtctttttt ccggctcagt catcgcccaa gctggcgcta tctgggcatc
     5161 ggggaggaag aagcccgtgc cttttcccgc gaggttgaag cggcatggaa agagtttgcc
     5221 gaggatgact gctgctgcat tgacgttgag cgaaaacgca cgtttaccat gatgattcgg
     5281 gaaggtgtgg ccatgcacgc ctttaacggt gaactgttcg ttcaggccac ctgggatacc
     5341 agttcgtcgc ggcttttccg gacacagttc cggatggtca gcccgaagcg catcagcaac
     5401 ccgaacaata ccggcgacag ccggaactgc cgtgccggtg tgcagattaa tgacagcggt
     5461 gcggcgctgg gatattacgt cagcgaggac gggtatcctg gctggatgcc gcagaaatgg
     5521 acatggatac cccgtgagtt acccggcggg cgcgcttggc gtaatcatgg tcatagctgt
     5581 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5641 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5701 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5761 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5821 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5881 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5941 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6001 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6061 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6121 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6181 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6241 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6301 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6361 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     6421 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6481 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6541 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6601 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6661 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6721 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6781 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6841 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6901 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6961 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7021 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7081 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7141 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7201 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7261 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7321 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7381 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7441 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7501 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7561 taagggcgac acggaaatgt tgaatactca t
//
