LOCUS       40924_24168        4660 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHC04, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4660)
  AUTHORS   Lee M-C., Chou H-H., Marx CJ.
  TITLE     Asymmetric, bi-modal tradeoffs during adaptation of Methylobacterium
            to distinct growth substrates
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4660)
  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 4660)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4660)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
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..4660
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(71..89)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(96..112)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      254..634
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             1002..1793
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SRLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVIHGDACLPNIMVENGRFSGFNDCGRLGVADRYQDIA
                     LDTRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1814..2671
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2845..3433
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    3721..3742
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3757..3787
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3795..3811
                     /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)."
     primer_bind     3819..3835
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3935..4651
                     /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"
ORIGIN
        1 aagggcgaat tctgcagata tccatcacac tggcggccgc tcgagcatgc atctagaggg
       61 cccaattcgc cctatagtga gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt
      121 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
      181 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
      241 aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
      301 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
      361 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
      421 gggttccgat ttagagcttt acggcacctc gaccgcaaaa aacttgattt gggtgatggt
      481 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
      541 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat cgcggtctat
      601 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
      661 taacaaattc agggcgcaag ggctgctaaa ggaaccggaa cacgtagaaa gccagtccgc
      721 agaaacggtg ctgaccccgg atgaatgtca gctactgggc tatctggaca agggaaaacg
      781 caagcgcaaa gagaaagcag gtagcttgca gtgggcttac atggcgatag ctagactggg
      841 cggttttatg gacagcaagc gaaccggaat tgccagctgg ggcgccctct ggtaaggttg
      901 ggaagccctg caaagtaaac tggatggctt tcttgccgcc aaggatctga tggcgcaggg
      961 gatcaagatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     1021 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     1081 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     1141 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     1201 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     1261 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcgcc
     1321 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     1381 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     1441 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     1501 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     1561 tccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     1621 acgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gatacccgtg
     1681 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     1741 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgaattg
     1801 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     1861 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     1921 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     1981 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtca
     2041 tacactatta tcccgtattg acgccgggca agagcaactc ggtcgccggg cgcggtattc
     2101 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     2161 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     2221 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     2281 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagag
     2341 tgacaccacg atgcctgtag caatgccaac aacgttgcgc aaactattaa ctggcgaact
     2401 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     2461 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     2521 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     2581 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     2641 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     2701 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     2761 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     2821 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2881 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2941 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     3001 gtagccgtag ttagcccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3061 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3121 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3181 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagcattg
     3241 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3301 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3361 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3421 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3481 ttttgctcac atgttctttc ctgccttatc ccctgattct gtggataacc gtattaccgc
     3541 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3601 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     3661 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     3721 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     3781 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     3841 ttacgccaag cttggtaccg agctcggatc cactagtaac ggccgccagt gtgctgaatt
     3901 cgcccttggt acctctagaa gggagagacc ccgaatggtg agcaagggcg aggagctgtt
     3961 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     4021 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctgatctg
     4081 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcct
     4141 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     4201 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     4261 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     4321 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     4381 caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg
     4441 ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat
     4501 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccgccctgag
     4561 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     4621 gatcactctc ggcatggacg agctgtacaa gtaaagatct
//
