LOCUS       62056_12330        8184 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 8184)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8184
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        128..156
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             2520..3236
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             3261..3278
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     rep_origin      complement(3419..4007)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4181..5038)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5039..5143)
                     /label=AmpR promoter
     CDS             complement(5310..6683)
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="VNTSYSQSNLRHNQILIWLCILSFFSVLNEMVLNVSLPDIANDFN
                     KPPASTNWVNTAFMLTFSIGTAVYGKLSDQLGIKRLLLFGIIINCFGSVIGFVGHSFFS
                     LLIMARFIQGAGAAAFPALVMVVVARYIPKENRGKAFGLIGSIVAMGEGVGPAIGGMIA
                     HYIHWSYLLLIPMITIITVPFLMKLLKKEVRIKGHFDIKGIILMSVGIVFFMLFTTSYS
                     ISFLIVSVLSFLIFVKHIRKVTDPFVDPGLGKNIPFMIGVLCGGIIFGTVAGFVSMVPY
                     MMKDVHQLSTAEIGSVIIFPGTMSVIIFGYIGGILVDRRGPLYVLNIGVTFLSVSFLTA
                     SFLLETTSWFMTIIIVFVLGGLSFTKTVISTIVSSSLKQQEAGAGMSLLNFTSFLSEGT
                     GIAIVGGLLSIPLLDQRLLPMEVDQSTYLYSNLLLLFSGIIVISWLVTLNVYKHSQRDF
                     "
     CDS             complement(6886..7887)
                     /codon_start=1
                     /label=repB
                     /note="RepB replication protein"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
ORIGIN
        1 ttttaggttc taaatcgtgt ttttcttgga attgtgctgt tttatccttt accttgtcta
       61 caaacccctt aaaaacgttt ttaaaggctt ttaagccgtc tgtacgttcc ttaaggaatt
      121 ctcatgtttg acagcttatc atcgataagc tttaatgcgg tagtttatca cagttaaatt
      181 gctaacgcag tcaggcaccg tgtatgaaat ctaacaatgc gctcatcgtc atcctcggca
      241 ccgtcaccct ggatgctgta ggcataggct tggttatgcc ggtactgccg ggcctcttgc
      301 gggatatcgt ccattccgac agcatcgcca gtcactatgg cgtgctgcta gcgctatatg
      361 cgttgatgca atttctatgc gcacccgttc tcggagcact gtccgaccgc tttggccgcc
      421 gcccagtcct gctcgcttcg ctacttggag ccactatcga ctacgcgatc atggcgacca
      481 cacccgtcct gtggatctga cgcgtgtaac tgcgaaggta aaacgggtga ttgaaaataa
      541 actaacaaat gaagaggaac gaaagagaag tctagaattt gttacgttta tatcggatga
      601 attactgcaa aatgatacgg agcctcgcac atttattatt cagaactctc ttttatcgct
      661 agagaaaatc catactctcg cacagaccaa agaggtagag gaatataaag aagtcataac
      721 tactctgaca agacatgatt cataatttga aaagcaggta aactaaccca aaaggcaaga
      781 ctctccgtag tttagaaaag agcaggttgc taataacata taaacagcca gttgccgtta
      841 tgataggtga ctggctgcat gggatgaaaa ggtgagggtg gagacagaca taacactctt
      901 aatagaagag ggtaattctt tctcttttat agaaaatcaa ttaattgaaa gtagctcctt
      961 cattcttaag atcaacgtga tataggtttg ctaacctttg cgttcactta actaacttat
     1021 aggggtaaca cttaaaaaag aatcaataac gatagaaacc gctcctaaag caggtgcatt
     1081 ttttcctaac gaagaaggca atagttcaca tttattgtct aaatgagaat ggactctaga
     1141 agaaacttcg tttttaatcg tatttaaaac aatgggatga gattcaatta tatgatttct
     1201 caagataaca gcttctatat caaatgtatt aaggatattg gttaatccaa ttccgatata
     1261 aaagccaaag ttttgaagtg catttaacat ttctacatca tttttatttg cgcgttccac
     1321 aatctctttt cgagaaatat tcttttcttc tttagagagc gaagccagta acgctttttc
     1381 agaagcatat aattcccaac agcctcgatt tccacagctg catttgggtc cattaaaatc
     1441 tatcgtcata tgacccattt ccccagaaaa accctgaaca cctttataca attcgttgtt
     1501 aataacaagt ccagttccaa ttccgatatt aatactgatg taaacgatgt tttcatagtt
     1561 ttttgtcata ccaaatactt tttcaccgta tgctcctgca ttagcttcat tttcaacaaa
     1621 aaccggaaca ttaaactcac tctcaattaa aaactgcaaa tctttgatat tccaatttaa
     1681 gttaggcatg aaaataattt gctgatgacg atctacaagg cctggaacac aaattcctat
     1741 tccgactaga ccataagggg actcaggcat atgggttaca aaaccatgaa taagtgcaaa
     1801 taaaatctct tttacttcac tagcggaaga actagacaag tcagaagtct tctcgagaat
     1861 aatatttcct tctaagtcgg ttagaattcc gttaagatag tcgactccta tatcaatacc
     1921 aatcgagtag cctgcattct tattaaaaac aagcattaca ggtcttctgc cgcctctaga
     1981 ttgccctgcc ccaatttcaa aaataaaatc tttttcaagc agtgtattta cttgagagga
     2041 gacagtagac ttgtttaatc ctgtaatctc agagagagtt gccctggaga caggggagtt
     2101 cttcaaaatt tcatctaata ttaatttttg attcattttt tttactaaag cttgatctgc
     2161 aatttgaata ataaccactc ctttgtttat ccaccgaact aagttggtgt tttttgaagc
     2221 ttgaattaga tatttaaaag tatcatatct aatattataa ctaaattttc taaaaaaaac
     2281 attgaaataa acatttattt tgtatatgat gagataaagt tagtttattg gataaacaaa
     2341 ctaactcaat taagatagtt gatggataaa cttgttcact taaatcaaag ggggaaatgt
     2401 acaatgaaaa aagtacttat ggcattcatt atttgtttat cgctgattct atctgtttta
     2461 gccgctccgc cgtctggcgc aggcgccgca ttgaagatct ccggagctcc cgggatccga
     2521 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     2581 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     2641 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     2701 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     2761 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     2821 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     2881 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     2941 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     3001 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     3061 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     3121 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     3181 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtcgc
     3241 atgccggcct gagaggatcg catcaccatc accatcacta accggtccaa gaattggagc
     3301 caatcaattc ttgcggagaa ctgtgaatgc gcaaaccaac ccttggcaga acatatccat
     3361 cgcgtccgcc atctccagca gccgcacgcg gcgcatctcg ggccgcgttg ctggcgtttt
     3421 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3481 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3541 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3601 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3661 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3721 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3781 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3841 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3901 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3961 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4021 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4081 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4141 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4201 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4261 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4321 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4381 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4441 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca
     4501 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4561 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4621 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4681 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4741 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaacacggg
     4801 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4861 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4921 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4981 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5041 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5101 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5161 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     5221 tcacgaggcc ctttcgtctt caagaattcc tgttataaaa aaaggatcaa ttttgaactc
     5281 tctcccaaag ttgatccctt aacgatttag aaatcccttt gagaatgttt atatacattc
     5341 aaggtaacca gccaactaat gacaatgatt cctgaaaaaa gtaataacaa attactatac
     5401 agataagttg actgatcaac ttccataggt aacaaccttt gatcaagtaa gggtatggat
     5461 aataaaccac ctacaattgc aatacctgtt ccctctgata aaaagctggt aaagttaagc
     5521 aaactcattc cagcaccagc ttcctgctgt ttcaagctac ttgaaacaat tgttgatata
     5581 actgttttgg tgaacgaaag cccacctaaa acaaatacga ttataattgt catgaaccat
     5641 gatgttgttt ctaaaagaaa ggaagcagtt aaaaagctaa cagaaagaaa tgtaactccg
     5701 atgtttaaca cgtataaagg acctcttcta tcaacaagta tcccaccaat gtagccgaaa
     5761 ataatgacac tcattgttcc agggaaaata attacacttc cgatttcggc agtacttagc
     5821 tggtgaacat ctttcatcat ataaggaacc atagagacaa accctgctac tgttccaaat
     5881 ataattcccc cacaaagaac tccaatcata aaaggtatat ttttccctaa tccgggatca
     5941 acaaaaggat ctgttacttt cctgatatgt tttacaaata tcaggaatga cagcacgcta
     6001 acgataagaa aagaaatgct atatgatgtt gtaaacaaca taaaaaatac aatgcctaca
     6061 gacattagta taattccttt gatatcaaaa tgacctttta tccttacttc tttctttaat
     6121 aatttcataa gaaacggaac agtgataatt gttatcatag gaatgagtag aagataggac
     6181 caatgaatat aatgggctat cattccacca atcgctggac cgactccttc tcccatggct
     6241 actatcgatc caataagacc aaatgcttta cccctatttt cctttggaat atagcgcgca
     6301 actacaacca ttacgagtgc tggaaatgca gctgcaccag ccccttgaat aaaacgagcc
     6361 ataataagta aggaaaagaa agaatggcca acaaacccaa ttaccgaccc gaaacaattt
     6421 attataattc caaataggag taaccttttg atgcctaatt gatcagatag ctttccatat
     6481 acagctgttc caatggaaaa ggttaacata aaggctgtgt tcacccagtt tgtactcgca
     6541 ggtggtttat taaaatcatt tgcaatatca ggtaatgaga cgttcaaaac catttcattt
     6601 aatacgctaa aaaaagataa aatgcaaagc caaattaaaa tttggttgtg tcgtaaattc
     6661 gattgtgaat aggatgtatt cacatttcac cctccaataa tgagggcaga cgtagtttat
     6721 agggttaatg atacgcttcc ctcttttaat tgaaccctgt tacattcatt acacttcata
     6781 attaattcct cctaaacttg attaaaacat tttaccacat ataaactaag ttttaaattc
     6841 agtatttcat cacttataca acaatatggc ccgtttgttg aactactctt taataaaata
     6901 atttttccgt tcccaattcc acattgcaat aatagaaaat ccatcttcat cggctttttc
     6961 gtcatcatct gtatgaatca aatcgccttc ttctgtgtca tcaaggttta attttttatg
     7021 tatttctttt aacaaaccac cataggagat taacctttta cggtgtaaac cttcctccaa
     7081 atcagacaaa cgtttcaaat tcttttcttc atcatcggtc ataaaatccg tatcctttac
     7141 aggatatttt gcagtttcgt caattgccga ttgtatatcc gatttatatt tatttttcgg
     7201 tcgaatcatt tgaactttta catttggatc atagtctaat ttcattgcct ttttccaaaa
     7261 ttgaatccat tgtttttgat tcacgtagtt ttctgtattc ttaaaataag ttggttccac
     7321 acataccaat acatgcatgt gctgattata agaattatct ttattattta ttgtcacttc
     7381 cgttgcacgc ataaaaccaa caagattttt attaattttt ttatattgca tcattcggcg
     7441 aaatccttga gccatatctg acaaactctt atttaattct tcgccatcat aaacattttt
     7501 aactgttaat gtgagaaaca accaacgaac tgttggcttt tgtttaataa cttcagcaac
     7561 aaccttttgt gactgaatgc catgtttcat tgctctcctc cagttgcaca ttggacaaag
     7621 cctggattta caaaaccaca ctcgatacaa ctttctttcg cctgtttcac gattttgttt
     7681 atactctaat atttcagcac aatcttttac tctttcagcc tttttaaatt caagaatatg
     7741 cagaagttca aagtaatcaa cattagcgat tttcttttct ctccatggtc tcacttttcc
     7801 actttttgtc ttgtccacta aaacccttga tttttcatct gaataaatgc tactattagg
     7861 acacataata ttaaaagaaa cccccatcta tttagttatt tgtttggtca cttataactt
     7921 taacagatgg ggtttttctg tgcaaccaat tttaagggtt ttcaatactt taaaacacat
     7981 acataccaac acttcaacgc acctttcagc aactaaaata aaaatgacgt tatttctata
     8041 tgtatcaaga taagaaagaa caagttcaaa accatcaaaa aaagacacct tttcaggtgc
     8101 tttttttatt ttataaactc attccctgat ctcgacttcg ttcttttttt acctctcggt
     8161 tatgagttag ttcaaattcg ttct
//
