LOCUS       Exported                9185 bp ds-DNA     circular SYN 05-MAR-2019
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    pNOC-RLUC.dna
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9185)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 5, 2019 from SnapGene 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9185
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
     promoter        1..798
                     /label=LDSP promoter
                     /note="color: #ffffff"
     CDS             799..1824
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /note="color: #993366"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     terminator      1842..2018
                     /label=35S terminator
                     /note="color: #ffffff"
     protein_bind    2068..2192
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
                     /note="color: #31849b"
     promoter        2217..2247
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
                     /note="color: #ffffff"
     CDS             2301..2960
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /note="color: #993366"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             3302..3607
                     /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."
                     /note="color: #993366"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(3648..3772)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
                     /note="color: #31849b; direction: LEFT"
     CDS             3792..4727
                     /codon_start=1
                     /label=Rlux
                     /note="color: #993366"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNDT"
     terminator      4734..6401
                     /label=LDSP terminator
                     /note="color: #ffffff"
     rep_origin      complement(6860..7448)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(7619..8479)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="color: #993366"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8480..8584)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      complement(8662..9117)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
ORIGIN
        1 gtaggtctct aagatggagt ggatggagga ggaggcgagc gtagcagcaa gcgtgagtta
       61 tacagccagg cacatgtcgc aatccttcgg tctcgggctt aaaatccacg cactaatcac
      121 gctgggccat gcaaagagca atgccgaggc ccaccacaca aaacgctgtg tcgcgcgttg
      181 cggcctgaag cttcatactt cttagtcgcc gccaaaaggg ctcgagagac gagacccgtt
      241 ggcatgaccg atgttgttcg acgcggtttg cttcgtcaca gtcgacgtga ttcaggaatc
      301 tggagcctgc agatcatttt tttcagcctg atatcgttct tttccactga gaaccatcag
      361 accacctttt cttccattgt gtgaaggagt aggagttgcc gtgctgcttt gtgggagaca
      421 tctgcgatgg tgaccagcct cccgtcgtct ggtcgacgtg acgagcctct tcactgttct
      481 tcgacggaga gacgcaagcg agacggctct agaccttttg gacacgcatt ctgtgtgtga
      541 actagtggac agtgatacca cgtctgaaag ctcaccactg cccatggtgc agctacttgt
      601 cacaaagttt tgactccgtc ggtatcacca ttcgcgctcg tgtgcctggt tgttccgcca
      661 cgccggcctg ccccggggcg gggcaatatt ctaaaatctc acgcaaaaca ccgcacttac
      721 ccctcacaca tattcgtgat agaccaccac caatctcagc ccgcatcaac acaggagggc
      781 cccggggggc aataagatat gaaaaagcct gaactcaccg cgacgtctgt cgagaagttt
      841 ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct
      901 cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc
      961 gatggtttct acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt
     1021 ccggaagtgc ttgacattgg ggaattcagc gagagcctga cctattgcat ctcccgccgt
     1081 gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg
     1141 gtcgcggagg ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc
     1201 ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt
     1261 gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc
     1321 gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc
     1381 gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc
     1441 attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc
     1501 tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg
     1561 gagcttgcag gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc
     1621 tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac
     1681 gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg
     1741 gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc
     1801 actcgtccga gggcaaagga atagagtaga tgccgaccgg agtccgcaaa aatcaccagt
     1861 ctctctctac aaatctatct ctctctattt ttctccagaa taatgtgtga gtagttccca
     1921 gataagggaa ttagggttct tatagggttt cgctcatgtg ttgagcatat aagaaaccct
     1981 tagtatgtat ttgtatttgt aaaatacttc tatcaataaa atttctaatt cctaaaacca
     2041 aaatccagtg acctgcaggc gatatcaaca agtttgtaca aaaaagctga acgagaaacg
     2101 taaaatgata taaatatcaa tatattaaat tagattttgc ataaaaaaca gactacataa
     2161 tactgtaaaa cacaacatat ccagtcacta tggcggccgc attaggcacc ccaggcttta
     2221 cactttatgc ttccggctcg tataatgtgt ggattttgag ttaggatccg gcgagatttt
     2281 caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat
     2341 cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata
     2401 accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca
     2461 agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaattcc
     2521 gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg
     2581 ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc
     2641 ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt
     2701 tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca
     2761 ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg
     2821 gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg
     2881 ccgtctgtga tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg
     2941 agtggcaggg cggggcgtaa agatctggat ccggcttact aaaagccaga taacagtatg
     3001 cgtatttgcg cgctgatttt tgcggtataa gaatatatac tgatatgtat acccgaagta
     3061 tgtcaaaaag aggtgtgcta tgaagcagcg tattacagtg acagttgaca gcgacagcta
     3121 tcagttgctc aaggcatata tgatgtcaat atctccggtc tggtaagcac aaccatgcag
     3181 aatgaagccc gtcgtctgcg tgccgaacgc tggaaagcgg aaaatcagga agggatggct
     3241 gaggtcgccc ggtttattga aatgaacggc tcttttgctg acgagaacag ggactggtga
     3301 aatgcagttt aaggtttaca cctataaaag agagagccgt tatcgtctgt ttgtggatgt
     3361 acagagtgat attattgaca cgcccgggcg acggatggtg atccccctgg ccagtgcacg
     3421 tctgctgtca gataaagtct cccgtgaact ttacccggtg gtgcatatcg gggatgaaag
     3481 ctggcgcatg atgaccaccg atatggccag tgtgccggtc tccgttatcg gggaagaagt
     3541 ggctgatctc agccaccgcg aaaatgacat caaaaacgcc attaacctga tgttctgggg
     3601 aatataaatg tcaggctccc ttatacacag ccagtctgca ggtcgaccat agtgactgga
     3661 tatgttgtgt tttacagtat tatgtagtct gttttttatg caaaatctaa tttaatatat
     3721 tgatatttat atcattttac gtttctcgtt cagctttctt gtacaaagtg gttgatagct
     3781 tggcgcgcct catggccagc aaggtgtacg accccgagca gcgcaagcgc atgatcaccg
     3841 gccctcagtg gtgggctcgc tgcaagcaga tgaacgtgct ggacagcttc atcaactact
     3901 acgacagcga gaagcacgcc gagaacgccg tgatcttcct gcacggcaac gccgccagca
     3961 gctacctgtg gcgccacgtg gtgccccaca tcgagcccgt ggcccgctgc atcatccccg
     4021 acctgatcgg catgggcaag agcggcaaga gcggcaacgg cagctaccgc ctgctggacc
     4081 actacaagta cctgaccgcc tggttcgagc tgctgaacct gcccaagaag atcatcttcg
     4141 tgggccacga ctggggcgcc tgcctggcct tccactacag ctacgagcac caggacaaga
     4201 tcaaggccat cgtgcacgcc gagagcgtgg tggacgtgat cgagagctgg gacgagtggc
     4261 ccgacatcga ggaggacatc gccctgatca agagcgagga gggcgagaag atggtgctgg
     4321 agaacaactt cttcgtggag accatgctgc ccagcaagat catgcgcaag ctggagcccg
     4381 aggagttcgc cgcctacctg gagcccttca aggagaaggg cgaggtgcgc cgtcccaccc
     4441 tgagctggcc tcgcgagatc cccctggtga agggcggcaa gcccgacgtg gtgcagatcg
     4501 tgcgcaacta caacgcctac ttgcgcgcca gcgacgacct gcccaagatg ttcatcgaga
     4561 gcgaccccgg cttcttcagc aacgccatcg tggagggcgc caagaagttc cccaacaccg
     4621 agttcgtgaa ggtgaagggc ctgcacttca gccaggagga cgctcccgac gagatgggca
     4681 agtacatcaa gagcttcgtg gagcgcgtgc tgaagaacga tacgtaagag ctcgaaagat
     4741 ccaagagaga cgagtagaga tttttttttt gggattgatg tttgtcgttc tttgagttgt
     4801 cgtcgagtta cgccttttgt aagaatgttc cgcaggagag gaggaggatg ggcatgagtg
     4861 agggtgagag ggcttgcccg cttttttttt taaaaacgct gctcttcgtg gttgtcaaac
     4921 aaacccccca tagaaacgat tttgttacgg tgcggtccag acgtcacttg aatggctccg
     4981 cggaaaggcc agggagggaa ggggggaggg aggaaacatg aaacatgttg aacggctcaa
     5041 cagggtttgg gggacaagag aggtagcgcc ctgatggact gctccctccc ctcctttccc
     5101 tcaatgtctc attcatccat gcttccccct tctctctctc ccctccgttc catcccccgc
     5161 gggcgtggta gtggcgtgat gggatccact aaaatgtacg tgtaagaaaa gccggtgagc
     5221 ttacgctttt gtgaaagtgg gagtacgagt gttgtgtgtg tgtgtagtgg tttcagaccc
     5281 cagacagagg cgaagcagaa aaagcagacg atgaagacga cgaagaaatg agcagtctat
     5341 ttttatcgtg gaaacagaag aggtgatatc gtctcgttct ttgttatcac ctaccccgcg
     5401 tgcatgtaca tgcagccttt ttattttgta atctttcccg aaaaatcaac cgccacctcc
     5461 cccccgcctt ctctcaccca tcatcttctc ctgtttatct tctactttac actagatcgc
     5521 atggcacatc tccctcgcaa tccatcggtg caaccatcat cgatcccact cctccctccc
     5581 tccctccctc cctcctcgca tctcttctaa gaaatccgct agctgcgaac gcagctcacc
     5641 tacctcattt atcagagcct cgattcggtc cttttccttc tcccctacct cgccgcaaac
     5701 agcaacagaa gcaggagcag caggagcagc ggcacctgtg tcaacagcac aagggcaagt
     5761 acagacagaa gacgaggcgg aggaggagga agacgatgtg atgtcattgg agctggctaa
     5821 actattagcc gttgctctca ccgcctttcc ccaaaactct tccacccatc gtgtcactcc
     5881 ttctatcagg ggaaccatcc ctcttttcct ccccaccttc ttccccttgc tcttcttctt
     5941 cctttgtctc tttgttcccc ctcctctctt cgcctcctcc tccgcatcgt cctctccttt
     6001 tatcaagtct tgcgttgcaa aggaagcaag ctctttgtga aggaccgcgc aagggatagg
     6061 agagtggaga gtgaggtgat gcagacaaac cgaggattcc cgggcgtcag ataacctaaa
     6121 atcgactact tcttgaggtg gaggaggagg gggaggagac ggagaataag tgtgtgtaga
     6181 aatatagctt gctgcctctg cttccatctt cgataaaatt gctgttgctg ttgctgttgc
     6241 ggctgatatc tgctgcggct gctgctgcac ctcctgaacg aagactgcag ccgcgtcttc
     6301 accctcttgg actactctgc cctcctccct tgcctttaca actcgatcga cgagattcgt
     6361 gccagtgctg tcatcatcat tggaaagagc aacagcatca ccttaagatt gaatcctgtt
     6421 gccggtcttg cgatgattat catataattt ctgttgaatt acgttaagca tgtaataatt
     6481 aacatgtaat gcatgacgtt atttatgaga tgggttttta tgattagagt cccgcaatta
     6541 tacatttaat acgcgataga aaacaaaata tagcgcgcaa actaggataa attatcgcgc
     6601 gcggtgtcat ctatgttact aggctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     6661 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6721 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6781 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6841 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6901 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6961 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     7021 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     7081 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     7141 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     7201 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     7261 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     7321 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7381 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     7441 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     7501 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7561 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7621 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7681 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7741 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7801 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7861 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7921 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7981 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     8041 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     8101 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     8161 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     8221 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8281 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     8341 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     8401 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     8461 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     8521 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     8581 cgcgcacatt tccccgaaaa gtgccacctg atgcggtgtg aaataccgca cagatgcgta
     8641 aggagaaaat accgcatcag gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     8701 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     8761 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     8821 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     8881 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     8941 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     9001 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     9061 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcca
     9121 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     9181 acgcc
//
