LOCUS       40924_14690        4762 bp DNA     circular SYN 18-DEC-2018
DEFINITION  E.coli-Thermotoga shuttle vector pDH10, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4762)
  AUTHORS   Han D, Norris SM, Xu Z.
  TITLE     Construction and transformation of a Thermotoga-E. coli shuttle 
            vector
  JOURNAL   BMC Biotechnol. 12, 2 (2012)
  PUBMED    22225774
REFERENCE   2  (bases 1 to 4762)
  AUTHORS   Han D, Xu Z.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-OCT-2011) Department of Biological Sciences, Bowling 
            Green State University, 217 Life Sci. Bldg., Bowling Green, OH 
            43403, USA
REFERENCE   3  (bases 1 to 4762)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4762)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "BMC 
            Biotechnol. 12, 2 (2012)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (04-OCT-2011) Department of Biological Sciences, Bowling Green State
            University, 217 Life Sci. Bldg., Bowling Green, OH 43403, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4762
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..125
                     /label=AmpR promoter
     CDS             126..983
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1157..1745
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2033..2054
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2069..2099
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2107..2123
                     /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     2131..2147
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2168..2186
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2216..2232
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             2440..2448
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             2458..3219
                     /codon_start=1
                     /gene="kan"
                     /product="thermostable kanamycin adenyltransferase"
                     /label=kan
                     /protein_id="AEW27249.1"
                     /translation="MNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGRQTD
                     GPYSDIEMMCVMSTEEAEFSHEWTTGEWKVEVNFYSEEILLDYASQVESDWPLTHGQFF
                     SILPIYDSGGYLEKVYQTAKSVEAQKFHDAICALIVEELFEYAGKWRNIRVQGPTTFLP
                     SLTVQVAMAGAMLIGLHHRICYTTSASLLTEAVKQSDLPSGYDHLCQFVMSGQLSDSEK
                     LLESLENFWNGIQEWTERHGYIVDVSKRIPF"
     gene            2458..3219
                     /gene="kan"
                     /label=kan
     CDS             complement(3229..3867)
                     /codon_start=1
                     /gene="rep"
                     /product="putative Rep protein"
                     /label=rep
                     /protein_id="AEW27248.1"
                     /translation="MSKEIITQNRVEQGSLDSKCHKSPLASTRRKKRFYQRAVSGVTAG
                     KHRNEFIAFLTLTSSLESPADITHSWEKLKKRIRRRYGNFEYIWVRERTQSGLVHMHIL
                     FRGPYIPQDWISKNWEEIHKAKIVYVEAVWDTGKAIRYMMKYLSKEMEGRFGYSWKWIF
                     KGAAQVWKWLCRALRYEMKEIIKIWEKLIIEIPPEGIRWGRIWELVGYG"
     gene            complement(3229..3867)
                     /gene="rep"
                     /label=rep
     promoter        complement(4119..4137)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4144..4160)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4302..4757
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
       61 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      121 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
      181 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
      241 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
      301 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
      361 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
      421 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
      481 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
      541 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
      601 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
      661 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
      721 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
      781 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
      841 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
      901 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
      961 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1021 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1081 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1141 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1201 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1261 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     1321 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1381 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1441 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1501 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     1561 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     1621 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     1681 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     1741 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     1801 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     1861 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     1921 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     1981 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     2041 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     2101 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     2161 agctcgaaat taaccctcac taaagggaac aaaagctggg taccgggccc cccctcgagg
     2221 tcgacggtat cgataagctt ggctgcaggt cgacggatcc ccgggagtat aacagaaacc
     2281 ttaaggcccg accgcttgac aagggcgcgt gaggttttta cgatagcgcc ggatgcgggg
     2341 aaaaagggct ccttttgggg ggttttcccc gcaccgggcg gacctgggcg cagaggaaac
     2401 gcggcaactc gcccgtctcg ggttcccgcc cacgaccctt aaggaggtgt gaggcatatg
     2461 aatggaccaa taataatgac tagagaagaa agaatgaaga ttgttcatga aattaaggaa
     2521 cgaatattgg ataaatatgg ggatgatgtt aaggctattg gtgtttatgg ctctcttggt
     2581 cgtcagactg atgggcccta ttcggatatt gagatgatgt gtgtcatgtc aacagaggaa
     2641 gcagagttca gccatgaatg gacaaccggt gagtggaagg tggaagtgaa tttttatagc
     2701 gaagagattc tactagatta tgcatctcag gtggaatcag attggccgct tacacatggt
     2761 caatttttct ctattttgcc gatttatgat tcaggtggat acttagagaa agtgtatcaa
     2821 actgctaaat cggtagaagc ccaaaagttc cacgatgcga tttgtgccct tatcgtagaa
     2881 gagctgtttg aatatgcagg caaatggcgt aatattcgtg tgcaaggacc gacaacattt
     2941 ctaccatcct tgactgtaca ggtagcaatg gcaggtgcca tgttgattgg tctgcatcat
     3001 cgcatctgtt atacgacgag cgcttcgctc ttaactgaag cagttaagca atcagatctt
     3061 ccttcaggtt atgaccatct gtgccagttc gtaatgtctg gtcaactttc cgactctgag
     3121 aaacttctgg aatcgctaga gaatttctgg aatgggattc aggagtggac agaacgacac
     3181 ggatatatag tggatgtgtc aaaacgcata ccattttgaa cggaattctt aaccatatcc
     3241 cactagttcc caaattcttc cccaccttat accctctggt ggtatctcaa taatcaactt
     3301 ctcccatatt ttgataatct ctttcatctc gtaccttaat gctctacata accacttcca
     3361 cacttgcgca gcccctttga aaatccactt ccaactatac ccgaatcttc cttccatctc
     3421 ctttgataag tacttcatca tatatcgtat tgctttacct gtgtcccata ctgcttctac
     3481 ataaacgatt tttgccttat gaatctcttc ccaattctta cttatccaat cttggggaat
     3541 ataaggtcct ctaaataata tatgcatatg caccaaacca ctttgagtac gttcccgtac
     3601 ccaaatgtat tcaaaattac catatcttct ccggattcgt tttttaagct tttcccagct
     3661 atgtgtgata tcggcaggag attccaatga agaggtaagc gtcaggaagg ctataaattc
     3721 gttcctatgc ttaccagcag ttacaccact tacagccctc tggtagaatc tcttttttcg
     3781 ccttgtagac gccagcggcg acttatgaca tttactatca agtgaacctt gctcgaccct
     3841 attttgtgtt ataatctctt ttgacatcgg gtagttacac cgcctttctt ttggattttt
     3901 ccccgggtgt atgtgggtac acccggggtt gtttcattat tatgagccct actaaatatt
     3961 ataagcacca tgtgttccct ataggtcaca ccataacctt atcttaatcc ttatcatctt
     4021 aagccttatc ataatcctta tcgtaaccct tatctaatca cactaaccac attcggatcc
     4081 tctagagcgg ccgccaccgc ggtggagctc caattcgccc tatagtgagt cgtattacaa
     4141 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     4201 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     4261 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
     4321 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4381 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4441 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4501 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4561 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4621 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4681 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4741 attaacgctt acaatttagg tg
//
