LOCUS       Exported                8440 bp ds-DNA     circular SYN 13-MAR-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8440)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 14, 2018 from SnapGene 4.0.5
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8440
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(323..341)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(348..364)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(505..960)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        987..1091
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1092..1952
                     /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      2123..2710
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2998..3019
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3034..3064
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3072..3088
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     3096..3112
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3133..3151
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3169..3732
                     /label=Ham34 promoter
     primer_bind     3745..3761
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3767..4483
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     terminator      4487..5007
                     /label=Ham34 terminator
     protein_bind    5183..5307
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(5348..5653)
                     /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
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(5995..6654)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(6708..6738)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(6764..6887)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        6902..7485
                     /label=Hsp70 promoter
     primer_bind     7069..7111
                     /label=Hsp70_IFF
     primer_bind     complement(7481..7520)
                     /label=Hsp70-GmR_R
     CDS             7501..8034
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     7501..7520
                     /label=GmR_F
     primer_bind     complement(8016..8065)
                     /label=Hsp70_IFR
     terminator      8221..8440
                     /label=Hsp70 terminator
ORIGIN
        1 tacgccgcct tagtaacaaa catgccaaaa gaaggaaagc tttcaccagc tttcaagttt
       61 aatcaatctt tgagagtcac aattgcaaga ataaatgaat ttttacaaca cagttcgatg
      121 cactgtgttt cgtcttgatg aaaattggac ggcaaatcga gtatcttaaa caaatctcgg
      181 cataggctgt ccattttgtg gtcgcctctt tgttggtagt ttgcagtagt ttcattagta
      241 gaagatctgc atcctcaatc tgctaaacgt cttgagcagc tatgagacga tccggggaat
      301 tggggcccga gctccaattc gccctatagt gagtcgtatt acaattcact ggccgtcgtt
      361 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
      421 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
      481 ttgcgcagcc tgaatggcga atggaaattg taagcgttaa tattttgtta aaattcgcgt
      541 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
      601 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
      661 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
      721 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
      781 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
      841 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
      901 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
      961 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     1021 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     1081 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     1141 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     1201 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     1261 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     1321 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     1381 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     1441 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     1501 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     1561 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     1621 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     1681 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     1741 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     1801 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     1861 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     1921 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     1981 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     2041 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     2101 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2161 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2221 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     2281 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2341 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     2401 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     2461 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     2521 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     2581 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     2641 tcgggtttcg ccacctctga cttgagcgtc gattttgtga tgctcgtcag gggggcggag
     2701 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     2761 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     2821 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     2881 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     2941 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     3001 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     3061 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     3121 cgccaagctc gaaattaacc ctcactaaag ggaacaaaag ctggtacctc tgatggacaa
     3181 agggtcgcct ctacgtaccc gcgacaatct cggcttttcc aatgtcgaca gctggcatat
     3241 cgcacgcgag gttaggcgtc ttcagccggc gtaaccacaa acctcgagtc aagtaccctt
     3301 tttcagacct taatttccgt gtccattccc tcgcgtttac acgaaggttt ctgagaaata
     3361 ccacatgtca cagagagaga gaagattgga gaaccttctc aacctgaccc gtgcagagcg
     3421 cttttgcgtc ctaccatccg ttaattatag caccaccacc acatgtctcg ccggaactct
     3481 cttatccgct ctttttgatc gtcttttttg cgccccatta aagacatctc ttgcacaggg
     3541 atcgtttctg attggccatt tcacttcctc ttttacatcc gaaaaccacc tgtttgtggg
     3601 gttgtgaagc cacaaccgga cgctcaaaaa gcctcgcccg actcgcccac gatcggaagg
     3661 ctcattctcc ttttcactct cacgtgccca agtcccaacc gactcttttt cgaccttcac
     3721 tctcaccgac aacgggcccc ccctcgaggt cgacggtatc gataccatgg tgagcaaggg
     3781 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     3841 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     3901 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt
     3961 cggctacggc ctgatgtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
     4021 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     4081 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     4141 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     4201 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     4261 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     4321 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca
     4381 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     4441 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaggccg aaggtcgagc
     4501 tgtgtgcact gagtgcgaag ttgcagattt tgtgtctatg catttaatac aatttccttt
     4561 tttccatttt cgttgttttt ttttgtgaaa gaacgattgt tttcaaaaga aggccgtttg
     4621 tagaacaatt tttttttata ctagcatttg gtattcgagt gtgacgaatc ggtgatggcg
     4681 ctttacaaat gaaatataac gctgaacaag cgcagtgaca gctttatttt ataacagaga
     4741 ataaagacca aagtagagta gactgatggc ggcttgaaac aaaactacgt gacttctaat
     4801 gcaaagaaaa ccatacccca cgctacttag aattgtttct ctttggttga tgaccgctat
     4861 agtcgtatgg agtgatccct ttttgtcaat atttcgattc ttgctaaaac cgtgactatt
     4921 ttgaggcaac tgtgtgtact taacatggta gaaatttcta tgtttaagcc agctgacaat
     4981 cattctttat ttatcgtaaa ccaaccaggt accgggccca ttatacctcg tattcgatta
     5041 aatgatacaa ctcattgcca caccgcaaga cggcgtcacc accatagaca gcctttcaag
     5101 cacccccttg tctatgagct gccacgtcag ctgccgctcc taatatccac ccgcgtccct
     5161 attcccgtca tatgccaagc tatcaacttt gtatagaaaa gttgaacgag aaacgtaaaa
     5221 tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta cataatactg
     5281 taaaacacaa catatccagt cactatggtc gacctgcaga ctggctgtgt ataagggagc
     5341 ctgacattta tattccccag aacatcaggt taatggcgtt tttgatgtca ttttcgcggt
     5401 ggctgagatc agccacttct tccccgataa cggagaccgg cacactggcc atatcggtgg
     5461 tcatcatgcg ccagctttca tccccgatat gcaccaccgg gtaaagttca cgggggactt
     5521 tatctgacag cagacgtgca ctggccaggg ggatcaccat ccgtcgcccg ggcgtgtcaa
     5581 taatatcact ctgtacatcc acaaacagac gataacggct ctctctttta taggtgtaaa
     5641 ccttaaactg catttcacca gcccctgttc tcgtcggcaa aagagccgtt catttcaata
     5701 aaccgggcga cctcagccat cccttcctga ttttccgctt tccagcgttc ggcacgcaga
     5761 cgacgggctt cattctgcat ggttgtgctt accgaaccgg agatattgac atcatatatg
     5821 ccttgagcaa ctgatagctg tcgctgtcaa ctgtcactgt aatacgctgc ttcatagcat
     5881 acctcttttt gacatacttc gggtatacat atcagtatat attcttatac cgcaaaaatc
     5941 agcgcgcaaa tacgcatact gttatctggc ttttagtaag ccggatcctc tagattacgc
     6001 cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa
     6061 gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg
     6121 cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt
     6181 taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat
     6241 aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat
     6301 gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt
     6361 ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc
     6421 tttcattgcc atacggaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa
     6481 ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag
     6541 ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt
     6601 acgatgccat tgggatatat caacggtggt atatccagtg atttttttct ccattttagc
     6661 ttccttagct cctgaaaatc tcgacggatc ctaactcaaa atccacacat tatacgagcc
     6721 ggaagcataa agtgtaaagc ctgggggtgc ctaatgcggc cgccatagtg actggatatg
     6781 ttgtgtttta cagtattatg tagtctgttt tttatgcaaa atctaattta atatattgat
     6841 atttatatca ttttacgttt ctcgttcaac tttattatac atagttgata attcacatat
     6901 gcgtcaatgc aattgactgt gtcctcgatt atttctatta ttaatattat ttccattaaa
     6961 aaagaaatcc agttcgtggc aaaataagac aaaggtttaa ataaaaattt atcgttatta
     7021 aactcgtaac ttaccttctt catggctcaa cccaatctgt atatcgactt atttaatttg
     7081 gttaacaaat cggttttcgt cgcaaatagg ggatgacaac taaaatacca atttgtgtcc
     7141 ttgcgacacg tcaacaggtg gctgccaata tctcaggttt tgagcacaca acgctgtgat
     7201 tggctatctg tagatcgctt cgagaacttt cggaccgaca tatcagaacg cagtcatgtc
     7261 caatccagct aacgattggt cgaaaccgca cttcatcgct tcatttcaac caattcccat
     7321 ctactaaaac gagcgtcgga atagtctcga acgtaacccc acgtagtggg tggcagcgcc
     7381 cctcgtatct ttcaatcgcc actcgagtct caagtttgct cactttgaaa ttttccatct
     7441 ggactacaca tcaaatctaa ctcaactttt atcaattgaa acacaagggc cccgtttcgc
     7501 atgttacgca gcagcaacga tgttacgcag cagggcagtc gccctaaaac aaagttaggt
     7561 ggctcaagta tgggcatcat tcgcacatgt aggctcggcc ctgaccaagt caaatccatg
     7621 cgggctgctc ttgatctttt cggtcgtgag ttcggagacg tagccaccta ctcccaacat
     7681 cagccggact ccgattacct cgggaacttg ctccgtagta agacattcat cgcgcttgct
     7741 gccttcgacc aagaagcggt tgttggcgct ctcgcggctt acgttctgcc caggtttgag
     7801 cagccgcgta gtgagatcta tatctatgat ctcgcagtct ccggcgagca ccggaggcag
     7861 ggcattgcca ccgcgctcat caatctcctc aagcatgagg ccaacgcgct tggtgcttat
     7921 gtgatctacg tgcaagcaga ttacggtgac gatcccgcag tggctctcta tacaaagttg
     7981 ggcatacggg aagaagtgat gcactttgat atcgacccaa gtaccgccac ctaagcggga
     8041 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     8101 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     8161 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccccg gatccccggg
     8221 gtcgactaga atacttagca attggttagt ttgaaaattg tcaaagcaga gttttttcga
     8281 caataaaatt tattttctaa atatttgatt tagcgttgac ttaatttgtt gattcggttg
     8341 tcatcttttg tgaagatgcc cgagagtcta gtaaatctca agcgaaacat agtgagaagg
     8401 tcactatggc tggtttgcat gagtctgggc ctattgtgct
//
