LOCUS       40924_24173        8828 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHC08, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8828)
  AUTHORS   Lee MC, Chou HH, Marx CJ.
  TITLE     Asymmetric, bimodal trade-offs during adaptation of Methylobacterium
            to distinct growth substrates
  JOURNAL   Evolution 63 (11), 2816-2830 (2009)
  PUBMED    19545267
REFERENCE   2  (bases 1 to 8828)
  AUTHORS   Chou H-H., Marx CJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-OCT-2008) Organismic and Evolutionary Biology, Harvard
            University, 16 Divinity Avenue, Biolab 3079, Cambridge, MA 02138, 
            USA
REFERENCE   3  (bases 1 to 8828)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8828)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Evolution";
            date: "2009"; volume: "63"; issue: "11"; pages: "2816-2830"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (14-OCT-2008) Organismic and Evolutionary Biology, Harvard 
            University, 16 Divinity Avenue, Biolab 3079, Cambridge, MA 02138, 
            USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    223..1120
                     /label=5' katA fragment
                     /note="5' katA fragment"
     misc_feature    1137..1170
                     /label=loxP site
                     /note="loxP site"
     protein_bind    1137..1170
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(1526..2173)
                     /codon_start=1
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     CDS             2279..3475
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="VKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     protein_bind    complement(3496..3529)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(3543..3559)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3783..3799
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      3981..4067
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4159..4186
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        4328..4356
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     CDS             4384..5091
                     /codon_start=1
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     terminator      5167..5214
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    5441..5806
                     /label=3' katA fragment
                     /note="3' katA fragment"
     primer_bind     complement(6607..6623)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6631..6647)
                     /label=lac operator
                     /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)."
     promoter        complement(6655..6685)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6700..6721)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7009..7597)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7771..8628)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8629..8733)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataaatt cgctcaccgc cggctcgcgt gggccggttc tgatgcagga ctaccacctc
      241 atcgagaaga tggcccactt caaccgggag cggatcccgg agcgggtggt gcacgccaag
      301 ggctacggcg cctacggcac gttccggctc accagaagcc tgtccgagtt cacccgtgcc
      361 aaggtgctga ccgaggtcgg caaggacgtg ccgatggtgg cgcgcttctc caccgtcggc
      421 ggcgagtccg gttcggccga caccgcccgc gacccgcgcg gcttcgccct caagttctac
      481 accgaggagg gcaactggga tctcgtcggc aacaacaccc cgatcttctt cgtgcgcgac
      541 gcgatcaagt tctccgactt catccgcacc cagaagcgcg atccgcgcac ccacctcaag
      601 ccgcattggc gccgctggga cttctggagc ctgtcgccgg aggcgatcca tcaggtcatg
      661 ttcctgtact cggatcgcgg cacgccgaaa tcggcgcgct tcatgaacgg ctacggctcg
      721 cacaccttct cgctctggaa cgaccggggc gagcgccact gggtgaagtt ccacttccac
      781 accaagcagg gcatccagaa cttctcggcc gacgaagccg acgaggtagc gggcaaggat
      841 ccggactact cctccgccga cctctcgact gccatcgagc agggcgactt cccgaaatgg
      901 aaggtctccg tgcagctcat gccggaattg gacgccgaca cctaccgcat caacccgttc
      961 gacctgacca aggtctggcc gcacggcgac tatccgctga tcgagattgg cgagatggag
     1021 ctcaaccgca atcccgagaa ctacttcgcc gagatcgagc aatctgcctt cgagccctcg
     1081 aacgtggtgc ccggcatcgg tttctcgccg cacaagatgc catgtatggc ggccgcataa
     1141 cttcgtatag catacattat acgaagttat ctagcgttca gggcatcaaa tgcagactcg
     1201 tttctgacac ttgcagactg gatctatctc atctgcgcaa ggcagaacgt gaagacggcc
     1261 gccctggacc tcgcccgcga gcgccaggcg cacgaggccg gcgcgcggac ccgcgccacg
     1321 gcccacgagc ggacgccgca gcaggagcgc cagaaggccg ccagagaggc cgagcgcggc
     1381 cgtgaggctt ggacgctagg gcagggcatg aaaaagcccg tagcgggctg ctacgggcgt
     1441 ctgacgcggt ggaaaggggg aggggatgtt gtctacatgg ctctgctgta gtgagtgggt
     1501 tgcgctccgg cagcggtcct gatcaatcgt caccctttct cggtccttca acgttcctga
     1561 caacgagcct ccttttcgcc aatccatcga caatcaccgc gagtccctgc tcgaacgctg
     1621 cgtccggacc ggcttcgtcg aaggcgtcta tcgcggcccg caacagcggc gagagcggag
     1681 cctgttcaac ggtgccgccg cgctcgccgg catcgctgtc gccggcctgc tcctcaagca
     1741 cggccccaac agtgaagtag ctgattgtca tcagcgcatt gacggcgtcc ccggccgaaa
     1801 aacccgcctc gcagaggaag cgaagctgcg cgtcggccgt ttccatctgc ggtgcgcccg
     1861 gtcgcgtgcc ggcatggatg cgcgcgccat cgcggtaggc gagcagcgcc tgcctgaagc
     1921 tgcgggcatt cccgatcaga aatgagcgcc agtcgtcgtc ggctctcggc accgaatgcg
     1981 tatgattctc cgccagcatg gcttcggcca gtgcgtcgag cagcgcccgc ttgttcctga
     2041 agtgccagta aagcgccggc tgctgaaccc ccaaccgttc cgccagtttg cgtgtcgtca
     2101 gaccgtctac gccgacctcg ttcaacaggt ccagggcggc acggatcact gtattcggct
     2161 gcaactttgt catgattgac actttatcac tgataaacat aatatgtcca ccaacttatc
     2221 agtgataaag aatccgcgcg ttcaatcgga ccagcggagg ctggtccgga ggccagacgt
     2281 gaaacccaac atacccctga tcgtaattct gagcactgtc gcgctcgacg ctgtcggcat
     2341 cggcctgatt atgccggtgc tgccgggcct cctgcgcgat ctggttcact cgaacgacgt
     2401 caccgcccac tatggcattc tgctggcgct gtatgcgttg gtgcaatttg cctgcgcacc
     2461 tgtgctgggc gcgctgtcgg atcgtttcgg gcggcggcca atcttgctcg tctcgctggc
     2521 cggcgccact gtcgactacg ccatcatggc gacagcgcct ttcctttggg ttctctatat
     2581 cgggcggatc gtggccggca tcaccggggc gactggggcg gtagccggcg cttatattgc
     2641 cgatatcact gatggcgatg agcgcgcgcg gcacttcggc ttcatgagcg cctgtttcgg
     2701 gttcgggatg gtcgcgggac ctgtgctcgg tgggctgatg ggcggtttct ccccccacgc
     2761 tccgttcttc gccgcggcag ccttgaacgg cctcaatttc ctgacgggct gtttcctttt
     2821 gccggagtcg cacaaaggcg aacgccggcc gttacgccgg gaggctctca acccgctcgc
     2881 ttcgttccgg tgggcccggg gcatgaccgt cgtcgccgcc ctgatggcgg tcttcttcat
     2941 catgcaactt gtcggacagg tgccggccgc gctttgggtc attttcggcg aggatcgctt
     3001 tcactgggac gcgaccacga tcggcatttc gcttgccgca tttggcattc tgcattcact
     3061 cgcccaggca atgatcaccg gccctgtagc cgcccggctc ggcgaaaggc gggcactcat
     3121 gctcggaatg attgccgacg gcacaggcta catcctgctt gccttcgcga cacggggatg
     3181 gatggcgttc ccgatcatgg tcctgcttgc ttcgggtggc atcggaatgc cggcgctgca
     3241 agcaatgttg tccaggcagg tggatgagga acgtcagggg cagctgcaag gctcactggc
     3301 ggcgctcacc agcctgacct cgatcgtcgg acccctcctc ttcacggcga tctatgcggc
     3361 ttctataaca acgtggaacg ggtgggcatg gattgcaggc gctgccctct acttgctctg
     3421 cctgccggcg ctgcgtcgcg ggctttggag cggcgcaggg caacgagccg atcgctgatc
     3481 gtggaaacga tctagataac ttcgtatagc atacattata cgaagttatg gatccagctt
     3541 atcgataccg tcgacctcga gaagaacaaa gaatacaaga aaatatttac aaaaaatcaa
     3601 tttaacaatt ccttaaaaca tgcaggaatt gacgatttaa acaatattag ctttgaacaa
     3661 ttcttatctc ttttcaatag ctataaatta tttaataagt aagttaaggg ataaactgca
     3721 tcccttaact tgtttttcgt gtgcctattt tttgtgaatc gatagatccc agtcacgacg
     3781 ttgtaaaacg acggccagtg aatttttcag cctgatacag attaaatcag aacgcagaag
     3841 cggtctgata aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat
     3901 gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag
     3961 agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     4021 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg
     4081 atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg
     4141 ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa
     4201 ctcttcctgt cgtcatatct acaagccatc cccccacaga tacggtaaac tagcctcgtt
     4261 tttgcatcag gaaagcaggg tcgacgcgtc tgcaggtcga ctctagtaag aaatctgaaa
     4321 tgagctgttg acaattaatc atcggctcgt ataatgtgtg gatctagaag ggagagaccc
     4381 cgaatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
     4441 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     4501 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
     4561 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     4621 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     4681 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     4741 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     4801 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     4861 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     4921 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     4981 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     5041 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaaagatct
     5101 cgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
     5161 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
     5221 ggaggaacta tatccggata tccacaggac gggtgtggtc gccatgatcg cgtagtcgat
     5281 agtggctcca agtagcgaag cgagcaggac tgggcggcgg ccaaagcggt cggacagtgc
     5341 tccgagaacg ggtgcgcata gaaattgcat caacgcatat agcgctagca gcacgccata
     5401 gtgactggcg atgctgtcgg aatggacgat atcgcctcgc gacggcgcca tgcgcaccga
     5461 cggcaatcac ggcgcccagg tcgattacga gccgaactcc ttcggcggtc cgaaacagga
     5521 cccgtccttc agcgagccgc cactgcggat ccagggcgat gctggccgct acggttggcc
     5581 gggcgacgac gaggatctct acggtcagcc caagctgttc tggaccaagg tgctggatga
     5641 gggcggccgc aagcggctcg tcgagaatat cgtcacctcg atgggcgaca gcccgcgcca
     5701 cattcaggag cggatgatcg cgcactggtt caaggtgcac gaggatttcg ggcgcggtgt
     5761 ggcccagggg ctcggcatcg atacggcaca ggccgccgcc gaatagtcat ctcaacatta
     5821 ttttgaatac aggggcatcg acgttggtcg gtgcctcttt tttgtgcgga cgaggcaaag
     5881 ccttgccttt tagaccactt ggaaggctgg aaacatcgtg cttatagcgt cggccaaggt
     5941 cgcgcggttc gtcacgcgcg tgccagaacc aacagggagt gccacgatgg ctgcaagcgc
     6001 agcaccggcc tggaccgggc agacggcgga agccaaggac ctttacgagc tcggcgagat
     6061 ccccccgctc ggccacgtgc ctgccaagat gtatgcctgg gcgatccgcc gcgagcgcca
     6121 cgggccgccg gagcagtcgc accagctcga agtgcttccc gtctgggaga tcggcgacga
     6181 cgaggtgctc gtctacgtca tggccgcggg cgtgaactac aacggcgttt gggccggcct
     6241 gggcgagcct atctcgccgt tcgacgtgca caagggcgag taccacatcg ccggctcgga
     6301 cgcgtcgggt atcgtctgga aggtcggcgc caaggtgaag cgctggaagg tcggcgacga
     6361 ggtcatcgtc cattgtaacc aggacgacgg cgacgacgag gagtgcaacg gcggcgatcc
     6421 gatgttctcg ccgacccagc ggatctgggg ttacgagacc ggcgacggtt cgttcgcgca
     6481 gttctgccgg gtgcagtcgc gtcagctcat ggcccgcccc aagcacctga cctgggaaga
     6541 ggccgcctgc tacacgctga cgctcgccac cgcctaccgc atgcaagcta gcttggcgta
     6601 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     6661 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     6721 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     6781 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     6841 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     6901 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     6961 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     7021 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     7081 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     7141 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     7201 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     7261 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     7321 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     7381 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     7441 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     7501 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     7561 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     7621 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7681 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     7741 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     7801 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     7861 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7921 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7981 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     8041 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     8101 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     8161 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     8221 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     8281 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     8341 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     8401 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     8461 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     8521 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     8581 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     8641 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     8701 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     8761 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     8821 ctttcgtc
//
