LOCUS       Exported                7690 bp ds-DNA     circular SYN 24-OCT-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7690)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Aug 12, 2020 from SnapGene Viewer 5.0.8
            https://www.snapgene.com
COMMENT     pDest-606 was digested in a 5ug reaction with SnaB1 and Xho1 in 
            Buffer 4 with BSA for 3 hours at 37degrees.
            This digest was cleaned and then 2ul was run on a gel to verify.  
            There is definitely a doublet of the upper band in this digest, 
            worse than the 605 digest showed.  However, I am hoping that this is
            just knciked, and will not interfere and come through the ligation.
            The digest was Klenow treated following the typical protocol and was
            then cleaned again.  This cleaned Klenow-treated vector was then 
            used in a ligation, 1ul of the vector and no insert piece in order 
            to allow for a religation of the backbone vector.  This ligation was
            Xfed into DB3.1 cells and plated on A/C at 37degrees. 1/12/2010  JM
            There were 12 colonies on the plate.  8 were picked. 1/13/2010  JM
            All 8 colonies are correct and show the smaller size that 
            corresponds to the vector without the GFP insert.  4 will be sent to
            sequencing to start with and the rest will be kept as back-up. 
            1/14/2010  JM
FEATURES             Location/Qualifiers
     source          1..7690
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..92
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     CDS             146..163
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             167..1264
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      1121..1130
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    1283..1407
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1432..1462
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1516..2175
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2517..2822
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2863..2987)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    3132..3266
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     mobile_element  3295..3460
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3644..4099
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4126..4230
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4231..5091
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5262..5850
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     mobile_element  6153..6377
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6444..6977)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(7166..7194)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        complement(7558..7667)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
ORIGIN
        1 atcatggaga taattaaaat gataaccatc tcgcaaataa ataagtattt tactgttttc
       61 gtaacagttt tgtaataaaa aaacctataa atattccgga ttattcatac cgtcccacca
      121 tcgggcgcgg atcaccatgg gatcccatca ccatcaccat cacggcaaaa tcgaagaagg
      181 taaactggta atctggatta acggcgataa aggctataac ggtctcgctg aagtcggtaa
      241 gaaattcgag aaagataccg gaattaaagt caccgttgag catccggata aactggaaga
      301 gaaattccca caggttgcgg caactggcga tggccctgac attatcttct gggcacacga
      361 ccgctttggt ggctacgctc aatctggcct gttggctgaa atcaccccgg acaaagcgtt
      421 ccaggacaag ctgtatccgt ttacctggga tgccgtacgt tacaacggca agctgattgc
      481 ttacccgatc gctgttgaag cgttatcgct gatttataac aaagatctgc tgccgaaccc
      541 gccaaaaacc tgggaagaga tcccggcgct ggataaagaa ctgaaagcga aaggtaagag
      601 cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg ccgctgattg ctgctgacgg
      661 gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt aaagacgtgg gcgtggataa
      721 cgctggcgcg aaagcgggtc tgaccttcct ggttgacctg attaaaaaca aacacatgaa
      781 tgcagacacc gattactcca tcgcagaagc tgcctttaat aaaggcgaaa cagcgatgac
      841 catcaacggc ccgtgggcat ggtccaacat cgacaccagc aaagtgaatt atggtgtaac
      901 ggtactgccg accttcaagg gtcaaccatc caaaccgttc gttggcgtgc tgagcgcagg
      961 tattaacgcc gccagtccga acaaagagct ggcaaaagag ttcctcgaaa actatctgct
     1021 gactgatgaa ggtctggaag cggttaataa agacaaaccg ctgggtgccg tagcgctgaa
     1081 gtcttacgag gaagagttgg cgaaagatcc acgtattgcc gccaccatgg aaaacgccca
     1141 gaaaggtgaa atcatgccga acatcccgca gatgtccgct ttctggtatg ccgtgcgtac
     1201 tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat gaagccctga aagacgcgca
     1261 gactaattcg ggatcctcga tcacaagttt gtacaaaaaa gctgaacgag aaacgtaaaa
     1321 tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta cataatactg
     1381 taaaacacaa catatccagt cactatggcg gccgcattag gcaccccagg ctttacactt
     1441 tatgcttccg gctcgtataa tgtgtggatt ttgagttagg atccggcgag attttcagga
     1501 gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa
     1561 tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag
     1621 accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt
     1681 tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga attccgtatg
     1741 gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc
     1801 catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag
     1861 tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct
     1921 aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt
     1981 tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa
     2041 tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtc
     2101 tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg
     2161 cagggcgggg cgtaaacgcg tggatccggc ttactaaaag ccagataaca gtatgcgtat
     2221 ttgcgcgctg atttttgcgg tataagaata tatactgata tgtatacccg aagtatgtca
     2281 aaaagaggtg tgctatgaag cagcgtatta cagtgacagt tgacagcgac agctatcagt
     2341 tgctcaaggc atatatgatg tcaatatctc cggtctggta agcacaacca tgcagaatga
     2401 agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat caggaaggga tggctgaggt
     2461 cgcccggttt attgaaatga acggctcttt tgctgacgag aacagggact ggtgaaatgc
     2521 agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga
     2581 gtgatattat tgacacgccc gggcgacgga tggtgatccc cctggccagt gcacgtctgc
     2641 tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc
     2701 gcatgatgac caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg
     2761 atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat
     2821 aaatgtcagg ctccgttata cacagccagt ctgcaggtcg accatagtga ctggatatgt
     2881 tgtgttttac agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata
     2941 tttatatcat tttacgtttc tcgttcagct ttcttgtaca aagtggtgat cgagctagct
     3001 agaagcttgt cgagaagtac tagaggatca taatcagcca taccacattt gtagaggttt
     3061 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     3121 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3181 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3241 tcaatgtatc ttatcatgtc tggatctgat cactgcttga gcctaggaga tccgaaccag
     3301 ataagtgaaa tctagttcca aactattttg tcatttttaa ttttcgtatt agcttacgac
     3361 gctacaccca gttcccatct attttgtcac tcttccctaa ataatcctta aaaactccat
     3421 ttccacccct cccagttccc aactattttg tccgcccaca gcggggcatt tttcttcctg
     3481 ttatgttttt aatcaaacat cctgccaact ccatgtgaca aaccgtcatc ttcggctact
     3541 ttttctctgt cacagaatga aaatttttct gtcatctctt cgttattaat gtttgtaatt
     3601 gactgaatat caacgcttat ttgcagcctg aatggcgaat gggacgcgcc ctgtagcggc
     3661 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3721 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3781 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3841 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3901 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3961 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     4021 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     4081 atattaacgt ttacaatttc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4141 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4201 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4261 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4321 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     4381 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4441 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     4501 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     4561 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     4621 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     4681 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     4741 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     4801 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4861 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4921 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4981 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5041 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5101 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5161 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5221 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5281 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5341 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5401 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5461 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5521 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5581 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5641 ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5701 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5761 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5821 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5881 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5941 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     6001 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     6061 catctgtgcg gtatttcaca ccgcagacca gccgcgtaac ctggcaaaat cggttacggt
     6121 tgagtaataa atggatgccc tgcgtaagcg ggtgtgggcg gacaataaag tcttaaactg
     6181 aacaaaatag atctaaacta tgacaataaa gtcttaaact agacagaata gttgtaaact
     6241 gaaatcagtc cagttatgct gtgaaaaagc atactggact tttgttatgg ctaaagcaaa
     6301 ctcttcattt tctgaagtgc aaattgcccg tcgtattaaa gaggggcgtg gccaagggca
     6361 tggtaaagac tatattcgcg gcgttgtgac aatttaccga acaactccgc ggccgggaag
     6421 ccgatctcgg cttgaacgaa ttgttaggtg gcggtacttg ggtcgatatc aaagtgcatc
     6481 acttcttccc gtatgcccaa ctttgtatag agagccactg cgggatcgtc accgtaatct
     6541 gcttgcacgt agatcacata agcaccaagc gcgttggcct catgcttgag gagattgatg
     6601 agcgcggtgg caatgccctg cctccggtgc tcgccggaga ctgcgagatc atagatatag
     6661 atctcactac gcggctgctc aaacctgggc agaacgtaag ccgcgagagc gccaacaacc
     6721 gcttcttggt cgaaggcagc aagcgcgatg aatgtcttac tacggagcaa gttcccgagg
     6781 taatcggagt ccggctgatg ttgggagtag gtggctacgt ctccgaactc acgaccgaaa
     6841 agatcaagag cagcccgcat ggatttgact tggtcagggc cgagcctaca tgtgcgaatg
     6901 atgcccatac ttgagccacc taactttgtt ttagggcgac tgccctgctg cgtaacatcg
     6961 ttgctgctgc gtaacatcgt tgctgctcca taacatcaaa catcgaccca cggcgtaacg
     7021 cgcttgctgc ttggatgccc gaggcataga ctgtacaaaa aaacagtcat aacaagccat
     7081 gaaaaccgcc actgcgccgt taccaccgct gcgttcggtc aaggttctgg accagttgcg
     7141 tgagcgcata cgctacttgc attacagttt acgaaccgaa caggcttatg tcaactgggt
     7201 tcgtgccttc atccgtttcc acggtgtgcg tcacccggca accttgggca gcagcgaagt
     7261 cgaggcattt ctgtcctggc tggcgaacga gcgcaaggtt tcggtctcca cgcatcgtca
     7321 ggcattggcg gccttgctgt tcttctacgg caaggtgctg tgcacggatc tgccctggct
     7381 tcaggagatc ggtagacctc ggccgtcgcg gcgcttgccg gtggtgctga ccccggatga
     7441 agtggttcgc atcctcggtt ttctggaagg cgagcatcgt ttgttcgccc aggactctag
     7501 ctatagttct agtggttggc ctactcgaga tcccgggtga tcaagtcttc gtcgagtgat
     7561 tgtaaataaa atgtaattta cagtatagta ttttaattaa tatacaaatg atttgataat
     7621 aattcttatt taactataat atattgtgtt gggttgaatt aaaggtccgt atactccgga
     7681 atattaatag
//
