LOCUS       62056_7625        9536 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 9536)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9536
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      102..556
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        582..686
                     /label=AmpR promoter
     CDS             687..1544
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1718..2306
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     mobile_element  complement(2611..2835)
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2905..3435)
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(3624..3652)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    complement(4169..4217)
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(4291..4998)
                     /codon_start=1
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     promoter        complement(5031..5140)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        5185..5276
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     enhancer        5457..5835
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5836..6039
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     intron          6175..6307
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     CDS             6419..6442
                     /codon_start=1
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
                     /translation="WSHPQFEK"
     CDS             6479..6502
                     /codon_start=1
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
                     /translation="WSHPQFEK"
     CDS             7565..7585
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     CDS             7604..8320
                     /codon_start=1
                     /label=mEGFP
                     /note="enhanced GFP with monomerizing A206K mutation
                     (Zacharias et al., 2002)"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             8321..8344
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     CDS             8345..8371
                     /codon_start=1
                     /label=9xHis
                     /note="9xHis affinity tag"
                     /translation="HHHHHHHHH"
     misc_feature    8405..8993
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    9126..9260
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     mobile_element  complement(9289..9454)
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 ttctctgtca cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga
       61 ctgaatatca acgcttattt gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
      121 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
      181 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
      241 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
      301 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
      361 ttttcgccct ttgacgttgg agtccacgtt cttaatagtg gactcttgtt ccaaactgga
      421 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
      481 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
      541 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
      601 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
      661 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
      721 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
      781 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
      841 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
      901 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
      961 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     1021 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     1081 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     1141 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     1201 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     1261 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     1321 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     1381 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     1441 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     1501 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     1561 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     1621 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     1681 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1741 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1801 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1861 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1921 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     1981 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2041 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2101 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2161 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2221 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2281 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     2341 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     2401 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     2461 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     2521 tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct ggcaaaatcg gttacggttg
     2581 agtaataaat ggatgccctg cgtaagcggg tgtgggcgga caataaagtc ttaaactgaa
     2641 caaaatagat ctaaactatg acaataaagt cttaaactag acagaatagt tgtaaactga
     2701 aatcagtcca gttatgctgt gaaaaagcat actggacttt tgttatggct aaagcaaact
     2761 cttcattttc tgaagtgcaa attgcccgtc gtattaaaga ggggcgtggc caagggcatg
     2821 gtaaagacta tattcgcggc gttgtgacaa tttaccgaac aactccgcgg ccgggaagcc
     2881 gatctcggct tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa agtgcatcac
     2941 ttcttcccgt atgcccaact ttgtatagag agccactgcg ggatcgtcac cgtaatctgc
     3001 ttgcacgtag atcacataag caccaagcgc gttggcctca tgcttgagga gattgatgag
     3061 cgcggtggca atgccctgcc tccggtgctc gccggagact gcgagatcat agatatagat
     3121 ctcactacgc ggctgctcaa acttgggcag aacgtaagcc gcgagagcgc caacaaccgc
     3181 ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta
     3241 atcggagtcc ggctgatgtt gggagtaggt ggctacgtct ccgaactcac gaccgaaaag
     3301 atcaagagca gcccgcatgg atttgacttg gtcagggccg agcctacatg tgcgaatgat
     3361 gcccatactt gagccaccta actttgtttt agggcgactg ccctgctgcg taacatcgtt
     3421 gctgctgcgt aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg
     3481 cttgctgctt ggatgcccga ggcatagact gtacaaaaaa acagtcataa caagccatga
     3541 aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg
     3601 agcgcatacg ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aactgggttc
     3661 gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg
     3721 aggcatttct gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg
     3781 cattggcggc cttgctgttc ttctacggca aggtgctgtg cacggatctg cccttgcttc
     3841 aggagatcgg tagacctcgg ccgtcgcggc gcttgccggt ggtgctgacc ccggatgaag
     3901 tggttcgcat cctcggtttt ctggaaggcg agcatcgttt gttcgcccag gactctagct
     3961 atagttctag tggttggcta cagctttgtt taaacaaagc tggctatggc agggcttgcc
     4021 gccccgacgt tggctgcgag ccctgggcct tcacccgaac ttgggggttg gggtggggaa
     4081 aaggaagaaa cgcgggcgta ttggtcccaa tggggtctcg gtggggtatc gacagagtgc
     4141 cagccctggg accgaacccc gcgtttatga acaaacgacc caacacccgt gcgttttatt
     4201 ctgtcttttt attgccgtca tagcgcgggt tccttccggt attgtctcct tccgtgtttc
     4261 agttagcctc ccccatctcc cggtacccta cttgtacagc tcgtccatgc cgccggtgga
     4321 gtggcggccc tcggcgcgtt cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt
     4381 gatgtccaac ttgatgttga cgttgtaggc gccgggcagc tgcacgggct tcttggcctt
     4441 gtaggtggtc ttgacctcag cgtcgtagtg gccgccgtcc ttcagcttca gcctctgctt
     4501 gatctcgccc ttcagggcgc cgtcctcggg gtacatccgc tcggaggagg cctcccagcc
     4561 catggtcttc ttctgcatta cggggccgtc ggaggggaag ttggtgccgc gcagcttcac
     4621 cttgtagatg aactcgccgt cctgcaggga ggagtcctgg gtcacggtca ccacgccgcc
     4681 gtcctcgaag ttcatcacgc gctcccactt gaagccctcg gggaaggaca gcttcaagta
     4741 gtcggggatg tcggcggggt gcttcacgta ggccttggag ccgtacatga actgagggga
     4801 caggatgtcc caggcgaagg gcagggggcc acccttggtc accttcagct tggcggtctg
     4861 ggtgccctcg taggggcggc cctcgccctc gccctcgatc tcgaactcgt ggccgttcac
     4921 ggagccctcc atgtgcacct tgaagcgcat gaactccttg atgatggcca tgttatcctc
     4981 ctcgcccttg ctcaccatgg tggccccggg tgatcaagtc ttcgtcgagt gattgtaaat
     5041 aaaatgtaat ttacagtata gtattttaat taatatacaa atgatttgat aataattctt
     5101 atttaactat aatatattgt gttgggttga attaaaggtc cgtatactag tatcgattcg
     5161 cgacctactc cggaatatta atagatcatg gagataatta aaatgataac catctcgcaa
     5221 ataaataagt attttactgt tttcgtaaca gttttgtaat aaaaaaacct ataaatattc
     5281 cggattattc ataccgtccc accatcgggc gcggatcctc aatattggcc attagccata
     5341 ttattcattg gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat
     5401 ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc atgttggcat
     5461 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     5521 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     5581 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     5641 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     5701 tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     5761 tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta cgtattagtc
     5821 atcgctatta ccatggtgat gcggttttgg cagtacacca atgggcgtgg atagcggttt
     5881 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     5941 caaaatcaac gggactttcc aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc
     6001 ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc
     6061 actagaagct ttattgcggt agtttatcac agttaaattg ctaacgcagt cagtgcttct
     6121 gacacaacag tctcgaactt aagctgcaga agttggtcgt gaggcactgg gcaggtaagt
     6181 atcaaggtta caagacaggt ttaaggagac caatagaaac tgggcttgtc gagacagaga
     6241 agactcttgc gtttctgata ggcacctatt ggtcttactg acatccactt tgcctttctc
     6301 tccacaggtg tccactccca gttcaattac agctcttaag ggatcccggt ccgaagcgcg
     6361 cgccaccatg aagaccatca tcgcactctc ttatattttc tgcctagtgt tcgcaggatg
     6421 gagccacccg cagttcgaga aaggtggagg ttcgggaggt ggatcgggtg gaagcgcttg
     6481 gagccacccg cagttcgaga aaggtggagg ttcgggaggt ggatcgggtg gaagcgctga
     6541 aaacctttac aacggcagcg aattcatgcc caatcagttc gtgcaaccag cctggcaaat
     6601 tgtcctttgg gcagctgcct acacggtcat tgtggtgacc tctgtggtgg gcaacgtggt
     6661 agtgatgtgg atcatcttag cccacaaaag aatgaggaca gtgacgaact attttctggt
     6721 gaacctggcc ttcgcggagg cctccatggc tgcattcaat acagtggtga acttcaccta
     6781 tgctgtccac aacgaatggt actacggcct gttctactgc aagttccaca acttctttcc
     6841 catcgccgct gtcttcgcca gtatctactc catgacggct gtggcctttg ataggtacat
     6901 ggccatcata catcccctcc agccccggct gtcagccaca gccaccaaag tggtcatctg
     6961 tgtcatctgg gtcctggctc tcctgctggc cttcccccag ggctactact caaccacaga
     7021 gaccatgccc agcagagtcg tgtgcatgat cgaatggcca gagcatccga acaagattta
     7081 tgagaaagtg taccacatct gtgtgactgt gctgatctac ttcctccccc tgctggtgat
     7141 tggctatgca tacaccgtag tgggaatcac actatgggcc agtgagatcc ccggggactc
     7201 ctctgaccgc taccacgagc aagtctctgc caagcgcaag gtggtcaaaa tgatgattgt
     7261 cgtggtgtgc accttcgcca tctgctggct gcccttccac atcttcttcc tcctgcccta
     7321 catcaaccca gatctctacc tgaagaagtt tatccagcag gtctacctgg ccatcatgtg
     7381 gctggccatg agctccacca tgtacaaccc catcatctac tgctgcctca atgacaggtt
     7441 ccgtctgggc ttcaagcatg ccttccggtg ctgccccttc atcagcgccg gcgactatga
     7501 ggggctggaa atgaaatcca cccggtatct ccagacccag ggcgcggccg ctttcgaatc
     7561 tagagaaaac ctttacttcc agggcaacgg cagcagcgga tctatggtga gcaagggcga
     7621 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     7681 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     7741 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     7801 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     7861 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     7921 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     7981 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     8041 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     8101 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     8161 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     8221 caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     8281 cgccgccggg atcactctcg gcatggacga gctgtacaag gactacaagg acgatgacga
     8341 caagcaccat catcaccatc atcaccacca ccactaagct agagcctgca gtctcgacaa
     8401 gcttaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     8461 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     8521 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     8581 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     8641 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     8701 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     8761 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     8821 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     8881 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     8941 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctgaagc
     9001 ttgtcgagaa gtactagagg atcataatca gccataccac atttgtagag gttttacttg
     9061 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     9121 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     9181 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     9241 tatcttatca tgtctggatc tgatcactgc ttgagcctag gagatccgaa ccagataagt
     9301 gaaatctagt tccaaactat tttgtcattt ttaattttcg tattagctta cgacgctaca
     9361 cccagttccc atctattttg tcactcttcc ctaaataatc cttaaaaact ccatttccac
     9421 ccctcccagt tcccaactat tttgtccgcc cacagcgggg catttttctt cctgttatgt
     9481 ttttaatcaa acatcctgcc aactccatgt gacaaaccgt catcttcggc tacttt
//
