LOCUS       40924_24153        8147 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHC01, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8147)
  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 8147)
  AUTHORS   Chou H-H., Marx CJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-OCT-2008) Organismic and Evolutionary Biology, Harvard
            University, 16 Divinity Avenue, Biolab 3079, Cambridge, MA 02138, 
            USA
REFERENCE   3  (bases 1 to 8147)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8147)
  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 
            (13-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..8147
                     /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"
     terminator      4486..4533
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    4760..5125
                     /label=3' katA fragment
                     /note="3' katA fragment"
     primer_bind     complement(5926..5942)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5950..5966)
                     /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(5974..6004)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6019..6040)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6328..6916)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7090..7947)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7948..8052)
                     /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 gaggcctcat atgtctagaa
     4381 ttcgagctcc cgggtaccat ggcatgcatc gatagatctc gagatccggc tgctaacaaa
     4441 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
     4501 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggatat
     4561 ccacaggacg ggtgtggtcg ccatgatcgc gtagtcgata gtggctccaa gtagcgaagc
     4621 gagcaggact gggcggcggc caaagcggtc ggacagtgct ccgagaacgg gtgcgcatag
     4681 aaattgcatc aacgcatata gcgctagcag cacgccatag tgactggcga tgctgtcgga
     4741 atggacgata tcgcctcgcg acggcgccat gcgcaccgac ggcaatcacg gcgcccaggt
     4801 cgattacgag ccgaactcct tcggcggtcc gaaacaggac ccgtccttca gcgagccgcc
     4861 actgcggatc cagggcgatg ctggccgcta cggttggccg ggcgacgacg aggatctcta
     4921 cggtcagccc aagctgttct ggaccaaggt gctggatgag ggcggccgca agcggctcgt
     4981 cgagaatatc gtcacctcga tgggcgacag cccgcgccac attcaggagc ggatgatcgc
     5041 gcactggttc aaggtgcacg aggatttcgg gcgcggtgtg gcccaggggc tcggcatcga
     5101 tacggcacag gccgccgccg aatagtcatc tcaacattat tttgaataca ggggcatcga
     5161 cgttggtcgg tgcctctttt ttgtgcggac gaggcaaagc cttgcctttt agaccacttg
     5221 gaaggctgga aacatcgtgc ttatagcgtc ggccaaggtc gcgcggttcg tcacgcgcgt
     5281 gccagaacca acagggagtg ccacgatggc tgcaagcgca gcaccggcct ggaccgggca
     5341 gacggcggaa gccaaggacc tttacgagct cggcgagatc cccccgctcg gccacgtgcc
     5401 tgccaagatg tatgcctggg cgatccgccg cgagcgccac gggccgccgg agcagtcgca
     5461 ccagctcgaa gtgcttcccg tctgggagat cggcgacgac gaggtgctcg tctacgtcat
     5521 ggccgcgggc gtgaactaca acggcgtttg ggccggcctg ggcgagccta tctcgccgtt
     5581 cgacgtgcac aagggcgagt accacatcgc cggctcggac gcgtcgggta tcgtctggaa
     5641 ggtcggcgcc aaggtgaagc gctggaaggt cggcgacgag gtcatcgtcc attgtaacca
     5701 ggacgacggc gacgacgagg agtgcaacgg cggcgatccg atgttctcgc cgacccagcg
     5761 gatctggggt tacgagaccg gcgacggttc gttcgcgcag ttctgccggg tgcagtcgcg
     5821 tcagctcatg gcccgcccca agcacctgac ctgggaagag gccgcctgct acacgctgac
     5881 gctcgccacc gcctaccgca tgcaagctag cttggcgtaa tcatggtcat agctgtttcc
     5941 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     6001 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6061 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6121 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6181 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6241 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6301 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6361 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6421 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6481 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6541 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6601 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6661 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6721 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6781 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6841 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6901 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6961 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7021 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7081 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7141 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7201 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7261 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7321 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7381 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7441 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7501 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7561 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7621 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7681 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7741 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7801 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7861 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7921 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7981 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     8041 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     8101 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtc
//
