LOCUS       62056_826        8826 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 8826)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8826
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2500..3210
                     /codon_start=1
                     /label=EYFP
                     /note="enhanced YFP"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     promoter        complement(4378..4396)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    4422..4646
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4692..5120
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5134..5464
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             5531..6322
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    6499..6632
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6669..6685)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6693..6709)
                     /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(6717..6747)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6762..6783)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7071..7659)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7833..8690)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8691..8795)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttaaccaa
      901 gaagctacta gaagaaatct tccctagatt cggcatgcca caggtattgg gaaccgacaa
      961 tgggcctgcc ttcgtctcca aggtaagtca gacagtagcc gatttattgg gggttgattg
     1021 gaaactacat tgtgcttaca gaccccagag ttcaggtcag gtagaaagaa tgaataggac
     1081 aatcaaggag actttaacta aattgacgct tgcaactggc tctagggact gggtgctcct
     1141 gcttcccctt gccctgtatc gagcccgcaa cacgccgggc ccccacggtc tcaccccata
     1201 tgaaatctta tatggggcac ccccgcccct tgtaaacttc cctgatcctg acatggcaaa
     1261 ggttactcat aacccctctc tccaagctca tttacaggca ctctacctgg tccagcacga
     1321 agtctggaga ccgttggcgg cagcttacca agagcaacta gaccggccgg tagtacctca
     1381 ccctttccgg gtcggtgaca cagtgtgggt ccgcagacac caaactaaaa atctagaacc
     1441 ccgctggaaa ggaccctata ccgtcctact gactaccccc accgctctca aagtagacgg
     1501 cattgcagcg tggatccacg ctgcccacgt aaaggctgcc gacaccagga ttgagccacc
     1561 agcagaatcg acatggcgtg ttcaacgctc ccaaaatccc ctaaagataa gattgacccg
     1621 cgggacctcc taatcccctt aattctcttc ctgtctctca aaggggccag atccgcagca
     1681 cccggctcca gccctcacca ggtctacaac attacctggg aagtgaccaa tggggatcgg
     1741 gagacagtat gggcaatatc aggcaaccac cctctgtgga cttggtggcc agtcctcacc
     1801 ccagatttgt gtatgttagc tctcagtggg ccgccccact gggggctaga gtatcaggcc
     1861 ccctattcct cgcccccggg gcccccttgt tgctcaggga gcagcgggag cagtgcaggc
     1921 tgttccagag actgcgacga gcccttgacc tccctcaccc ctcggtgcaa cactgcctgg
     1981 aacagactta agctagacca ggtaactcat aaatcaagtg agggatttta tgtctgcccc
     2041 gggtcacatc gcccccggga agccaagtcc tgtggaggtc cagactcctt ctactgtgcc
     2101 tcttggggct gcgagacaac cggtagagta tactggaagc cctcctcgag ttgggactac
     2161 atcacagtgg acaacaatct caccactagc caggctgtcc aggtatgcaa agacaataag
     2221 tggtgcaatc ccttggctat ccagtttaca aacgccggga aacaggtcac ctcatggaca
     2281 actggacact attggggtct acgtctttat gtctctgggc gggacccggg gcttactttc
     2341 gggatccgac tcagatatca aaatctagga cctcgggtac cgataggacc gaaccccgtc
     2401 ctggcagacc aactttcgct cccgcgacct aatcccctac ccaaacctgc caagtctccc
     2461 cccgcctcta attcgactcc cacattgatt tccgctagca gcaagggcga ggagctgttc
     2521 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     2581 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     2641 accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg
     2701 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     2761 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     2821 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     2881 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     2941 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     3001 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     3061 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc
     3121 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     3181 atcactctcg gcatggacga gctgtacaag gctagcgctg ttgccggcgc cccgtccccc
     3241 actcccactc agcccccgcc agcaggaacg ggagacaggt tactaaatct agtacaggga
     3301 gcttaccagg cactcaacct taccaaccct gataaaactc aagagtgctg gttatgccta
     3361 gtgtctggac ccccctatta cgaaggggtt gccgtcctag gtacttattc caaccatacc
     3421 tctgccccag ctaactgctc cgtggcctcc caacacaagt tgaccctgtc cgaagtgact
     3481 ggacggggac tctgcatagg aacagtccca aaaactcacc aggccctgtg caacactacc
     3541 cttaagatag acaaagggtc ttactatcta gttgccccca caggaactac gtgggcatgt
     3601 aacactggac tcactccatg cctatctgcc accgtgctta atcgcaccac tgactattgc
     3661 gttctcgtag agttatggcc cagggtcacc taccatcctc ccagttacgt ctatagccag
     3721 tttgaaaaat cctatagaca taaaagagaa ccagtgtcct taaccttggc cctattatta
     3781 ggtgggctaa ctatgggtgg catcgccgcg ggagtaggga caggaactac cgccctggtc
     3841 gccacccagc agttccagca gctccatgct gccgtacaag atgatctcaa agaagtcgaa
     3901 aagtcaatta ctaacctaga aaagtctctt acttcgttgt ctgaggttgt gctgcagaat
     3961 cgacgaggcc tagacctgtt gttcctaaaa gaaggaggac tgtgtgctgc cctaaaagaa
     4021 gaatgttgtt tctatgctga ccatacaggc ctagtaagag atagtatggc caaattaaga
     4081 gagagactca ctcagagaca aaaactattt gagtcgagcc aaggatggtt cgaaggattg
     4141 tttaacagat ccccctggtt taccacgtta atatccacca tcatggggcc tctcattata
     4201 ctcctactaa ttctgctttt tggaccctgc attcttaatc gattagtcca atttgttaaa
     4261 gacaggatat cagtggtcca ggctttagtt ttgactcaac aatatcacca gctgaagcca
     4321 ctagaatacg agccatgata agaattctgc agatatccat cacactggcg gccctattct
     4381 atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca
     4441 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     4501 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     4561 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     4621 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     4681 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4741 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4801 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     4861 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4921 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4981 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     5041 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     5101 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     5161 ccaggctccc caggcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     5221 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     5281 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     5341 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     5401 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     5461 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     5521 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     5581 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     5641 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     5701 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     5761 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     5821 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     5881 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     5941 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     6001 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     6061 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     6121 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     6181 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     6241 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     6301 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     6361 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     6421 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     6481 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     6541 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     6601 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     6661 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     6721 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6781 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6841 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6901 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6961 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     7021 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     7081 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     7141 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     7201 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     7261 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     7321 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     7381 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     7441 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     7501 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7561 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7621 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7681 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7741 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7801 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7861 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7921 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7981 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     8041 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     8101 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     8161 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     8221 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     8281 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     8341 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     8401 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     8461 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     8521 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     8581 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     8641 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     8701 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     8761 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     8821 gacgtc
//
