LOCUS       Exported                5522 bp ds-DNA     circular SYN 22-MAY-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5522)
  AUTHORS   Jianjian LI
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, May 23, 2019 from SnapGene 4.3.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5522
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        5..359
                     /label=CaMV 35S promoter
     primer_bind     291..310
                     /label=35S
     5'UTR           360..868
     protein_bind    874..998
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1035..1065
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1119..1778
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2120..2425
                     /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(2466..2590)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2613..2657
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2670..2714
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2727..2771
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2763..2791)
                     /label=AVI-R
     3'UTR           2792..2975
     terminator      3029..3281
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     rep_origin      complement(3519..4107)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4278..5138)
                     /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"
     promoter        complement(5139..5243)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tatgcggcca gtgaattgtt aattaagaat tcgagctcca ccgcggaaac ctcctcggat
       61 tccattgccc agctatctgt cactttattg agaagatagt ggaaaaggaa ggtggctcct
      121 acaaatgcca tcattgcgat aaaggaaagg ccatcgttga agatgcctct gccgacagtg
      181 gtcccaaaga tggaccccca cccacgagga gcatcgtgga aaaagaagac gttccaacca
      241 cgtcttcaaa gcaagtggat tgatgtgata tctccactga cgtaagggat gacgcacaat
      301 cccactatcc ttcgcaagac ccttcctcta tataaggaag ttcatttcat ttggagaggt
      361 attaaaatct taataggttt tgataaaagc gaacgtgggg aaacccgaac caaaccttct
      421 tctaaactct ctctcatctc tcttaaagca aacttctctc ttgtctttct tgcgtgagcg
      481 atcttcaacg ttgtcagatc gtgcttcggc accagtacaa cgttttcttt cactgaagcg
      541 aaatcaaaga tctctttgtg gacacgtagt gcggcgccat taaataacgt gtacttgtcc
      601 tattcttgtc ggtgtggtct tgggaaaaga aagcttgctg gaggctgctg ttcagcccca
      661 tacattactt gttacgattc tgctgacttt cggcgggtgc aatatctcta cttctgcttg
      721 acgaggtatt gttgcctgta cttctttctt cttcttcttg ctgattggtt ctataagaaa
      781 tctagtattt tctttgaaac agagttttcc cgtggttttc gaacttggag aaagattgtt
      841 aagcttctgt atattctgcc caaattcgat caaacaagtt tgtacaaaaa agctgaacga
      901 gaaacgtaaa atgatataaa tatcaatata ttaaattaga ttttgcataa aaaacagact
      961 acataatact gtaaaacaca acatatccag tcactatggc ggcccctcga ggggccgcat
     1021 taggcacccc aggctttaca ctttatgctt ccggctcgta taatgtgtgg attttgagtt
     1081 aggatccggc gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat
     1141 ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag
     1201 ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg
     1261 taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga
     1321 atgctcatcc ggaattccgt atggcaatga aagacggtga gctggtgata tgggatagtg
     1381 ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg
     1441 aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg
     1501 gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca
     1561 atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg
     1621 cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg
     1681 cgattcaggt tcatcatgcc gtctgtgatg gcttccatgt cggcagaatg cttaatgaat
     1741 tacaacagta ctgcgatgag tggcagggcg gggcgtaaag atctggatcc ggcttactaa
     1801 aagccagata acagtatgcg tatttgcgcg ctgatttttg cggtataaga atatatactg
     1861 atatgtatac ccgaagtatg tcaaaaagag gtgtgctatg aagcagcgta ttacagtgac
     1921 agttgacagc gacagctatc agttgctcaa ggcatatatg atgtcaatat ctccggtctg
     1981 gtaagcacaa ccatgcagaa tgaagcccgt cgtctgcgtg ccgaacgctg gaaagcggaa
     2041 aatcaggaag ggatggctga ggtcgcccgg tttattgaaa tgaacggctc ttttgctgac
     2101 gagaacaggg actggtgaaa tgcagtttaa ggtttacacc tataaaagag agagccgtta
     2161 tcgtctgttt gtggatgtac agagtgatat tattgacacg cccgggcgac ggatggtgat
     2221 ccccctggcc agtgcacgtc tgctgtcaga taaagtctcc cgtgaacttt acccggtggt
     2281 gcatatcggg gatgaaagct ggcgcatgat gaccaccgat atggccagtg tgccggtctc
     2341 cgttatcggg gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca aaaacgccat
     2401 taacctgatg ttctggggaa tataaatgtc aggctccctt atacacagcc agtctgcagg
     2461 tcgaccatag tgactggata tgttgtgttt tacagtatta tgtagtctgt tttttatgca
     2521 aaatctaatt taatatattg atatttatat cattttacgt ttctcgttca gctttcttgt
     2581 acaaagtggt tgggtaccct cgaggcggcc gcggcttgaa cgacattttc gaggcccaga
     2641 aaatcgagtg gcacgagggt gcgggcgctg ggctcaatga catctttgag gcgcaaaaaa
     2701 tagaatggca tgagggggct ggcgcgggcc ttaacgatat ttttgaagca caaaaaatcg
     2761 aatggcatga atagatatga agatgaagat gttaactctg gtttcattaa attttcttta
     2821 gtttgaattt actgttattc ggtgtgcatt tctatgtttg gtgagcggtt ttctgtgctc
     2881 agagtgtgtt tattttatgt aatttaattt ctttgtgagc tcctgtttag caggtcgtcc
     2941 cttcagcaag gacacaaaaa gattttaatt ttattaaaaa aaaaaaaaaa aaagaccggg
     3001 aattcgatat caagcttatc gacctgcaga tcgttcaaac atttggcaat aaagtttctt
     3061 aagattgaat cctgttgccg gtcttgcgat gattatcata taatttctgt tgaattacgt
     3121 taagcatgta ataattaaca tgtaatgcat gacgttattt atgagatggg tttttatgat
     3181 tagagtcccg caattataca tttaatacgc gatagaaaac aaaatatagc gcgcaaacta
     3241 ggataaatta tcgcgcgcgg tgtcatctat gttactagat ctctgcatta atgaatcggc
     3301 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     3361 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3421 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3481 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3541 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3601 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3661 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3721 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3781 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3841 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3901 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3961 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4021 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4081 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4141 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4201 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4261 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4321 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4381 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4441 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4501 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4561 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4621 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4681 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4741 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4801 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4861 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     4921 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4981 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5041 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5101 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5161 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5221 aataaacaaa tgggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     5281 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     5341 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     5401 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     5461 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     5521 ca
//
