LOCUS       40924_24163        5371 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHC03, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5371)
  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  JOURNAL   Unpublished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 5371)
  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 5371)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5371)
  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            "Unpublished"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..5371
                     /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..5362
                     /codon_start=1
                     /label=tdTomato
                     /note="tandem dimeric (pseudo-monomeric) derivative of
                     DsRed (Shaner et al., 2004)"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
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 aggaggtcat
     3961 caaagagttc atgcgcttca aggtgcgcat ggagggctcc atgaacggcc acgagttcga
     4021 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     4081 gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tccccccagt tcatgtacgg
     4141 ctccaaggcg tacgtgaagc accccgccga catccccgat tacaagaagc tgtccttccc
     4201 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggtctgg tgaccgtgac
     4261 ccaggactcc tccctgcagg acggcacgct gatctacaag gtgaagatgc gcggcaccaa
     4321 cttccccccc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccaccga
     4381 gcgcctgtac ccccgcgacg gcgtgctgaa gggcgagatc caccaggccc tgaagctgaa
     4441 ggacggcggc cactacctgg tggagttcaa gaccatctac atggccaaga agcccgtgca
     4501 actgcccggc tactactacg tggacaccaa gctggacatc acctcccaca acgaggacta
     4561 caccatcgtg gaacagtacg agcgctccga gggccgccac cacctgttcc tggggcatgg
     4621 caccggcagc accggcagcg gcagctccgg caccgcctcc tccgaggaca acaacatggc
     4681 cgtcatcaaa gagttcatgc gcttcaaggt gcgcatggag ggctccatga acggccacga
     4741 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     4801 gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc cccagttcat
     4861 gtacggctcc aaggcgtacg tgaagcaccc cgccgacatc cccgattaca agaagctgtc
     4921 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gtctggtgac
     4981 cgtgacccag gactcctccc tgcaggacgg cacgctgatc tacaaggtga agatgcgcgg
     5041 caccaacttc ccccccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     5101 caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccacc aggccctgaa
     5161 gctgaaggac ggcggccact acctggtgga gttcaagacc atctacatgg ccaagaagcc
     5221 cgtgcaactg cccggctact actacgtgga caccaagctg gacatcacct cccacaacga
     5281 ggactacacc atcgtggaac agtacgagcg ctccgagggc cgccaccacc tgttcctgta
     5341 cggcatggac gagctgtaca agtaaagatc t
//
