LOCUS       40924_24183        8837 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHC10, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8837)
  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 8837)
  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 8837)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8837)
  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..8837
                     /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..5100
                     /codon_start=1
                     /label=Venus
                     /note="yellow fluorescent protein (YFP) with fast and
                     efficient maturation (Nagai et al., 2002)"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     terminator      5176..5223
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    5450..5815
                     /label=3' katA fragment
                     /note="3' katA fragment"
     primer_bind     complement(6616..6632)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6640..6656)
                     /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(6664..6694)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6709..6730)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7018..7606)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7780..8637)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8638..8742)
                     /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 ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     4441 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     4501 tacggcaagc tgaccctgaa gctgatctgc accaccggca agctgcccgt gccctggccc
     4561 accctcgtga ccaccctggg ctacggcctg cagtgcttcg cccgctaccc cgaccacatg
     4621 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     4681 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     4741 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     4801 cacaagctgg agtacaacta caacagccac aacgtctata tcaccgccga caagcagaag
     4861 aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc
     4921 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     4981 cactacctga gctaccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     5041 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     5101 taaagatctc gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     5161 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     5221 ttgctgaaag gaggaactat atccggatat ccacaggacg ggtgtggtcg ccatgatcgc
     5281 gtagtcgata gtggctccaa gtagcgaagc gagcaggact gggcggcggc caaagcggtc
     5341 ggacagtgct ccgagaacgg gtgcgcatag aaattgcatc aacgcatata gcgctagcag
     5401 cacgccatag tgactggcga tgctgtcgga atggacgata tcgcctcgcg acggcgccat
     5461 gcgcaccgac ggcaatcacg gcgcccaggt cgattacgag ccgaactcct tcggcggtcc
     5521 gaaacaggac ccgtccttca gcgagccgcc actgcggatc cagggcgatg ctggccgcta
     5581 cggttggccg ggcgacgacg aggatctcta cggtcagccc aagctgttct ggaccaaggt
     5641 gctggatgag ggcggccgca agcggctcgt cgagaatatc gtcacctcga tgggcgacag
     5701 cccgcgccac attcaggagc ggatgatcgc gcactggttc aaggtgcacg aggatttcgg
     5761 gcgcggtgtg gcccaggggc tcggcatcga tacggcacag gccgccgccg aatagtcatc
     5821 tcaacattat tttgaataca ggggcatcga cgttggtcgg tgcctctttt ttgtgcggac
     5881 gaggcaaagc cttgcctttt agaccacttg gaaggctgga aacatcgtgc ttatagcgtc
     5941 ggccaaggtc gcgcggttcg tcacgcgcgt gccagaacca acagggagtg ccacgatggc
     6001 tgcaagcgca gcaccggcct ggaccgggca gacggcggaa gccaaggacc tttacgagct
     6061 cggcgagatc cccccgctcg gccacgtgcc tgccaagatg tatgcctggg cgatccgccg
     6121 cgagcgccac gggccgccgg agcagtcgca ccagctcgaa gtgcttcccg tctgggagat
     6181 cggcgacgac gaggtgctcg tctacgtcat ggccgcgggc gtgaactaca acggcgtttg
     6241 ggccggcctg ggcgagccta tctcgccgtt cgacgtgcac aagggcgagt accacatcgc
     6301 cggctcggac gcgtcgggta tcgtctggaa ggtcggcgcc aaggtgaagc gctggaaggt
     6361 cggcgacgag gtcatcgtcc attgtaacca ggacgacggc gacgacgagg agtgcaacgg
     6421 cggcgatccg atgttctcgc cgacccagcg gatctggggt tacgagaccg gcgacggttc
     6481 gttcgcgcag ttctgccggg tgcagtcgcg tcagctcatg gcccgcccca agcacctgac
     6541 ctgggaagag gccgcctgct acacgctgac gctcgccacc gcctaccgca tgcaagctag
     6601 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     6661 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     6721 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     6781 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     6841 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6901 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6961 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     7021 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     7081 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     7141 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     7201 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     7261 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     7321 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     7381 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     7441 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     7501 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     7561 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     7621 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     7681 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     7741 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     7801 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7861 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7921 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7981 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     8041 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     8101 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     8161 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     8221 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     8281 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     8341 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     8401 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     8461 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     8521 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     8581 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     8641 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     8701 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     8761 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     8821 cacgaggccc tttcgtc
//
