LOCUS       Exported                4550 bp ds-DNA     circular SYN 06-JUN-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4550)
  AUTHORS   Sean McGrath
  TITLE     Direct Submission
  JOURNAL   Exported Jun 6, 2018 from SnapGene 4.1.9
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4550
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        913..943
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             987..1057
                     /codon_start=1
                     /gene="lacZ (fragment)"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGDTIEYSSYASSLV"
     promoter        1009..1027
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     promoter        1078..1096
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1142..2059
                     /codon_start=1
                     /label=hNANOG
                     /translation="MSVDPACPQSLPCFEASDCKESSPMPVICGPEENYPSLQMSSAEM
                     PHTETVSPLPSSMDLLIQDSPDSSTSPKGKQPTSAEKSVAKKEDKVPVKKQKTRTVFSS
                     TQLCVLNDRFQRQKYLSLQQMQELSNILNLSYKQVKTWFQNQRMKSKRWQKNNWPKNSN
                     GVTQKASAPTYPSLYSSYHQGCLVNPTGNLPMWSNQTWNNSTWSNQTQNIQSWSNHSWN
                     TQTWCTQSWNNQAWNSPFYNCGEESLQSCMQFQPNSPASDLEAALEAAGEGLNVIQQTT
                     RYFSTPQTMDLFLNYSMNMQPEDV"
     CDS             2144..2377
                     /codon_start=1
                     /gene="lacZ (fragment)"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="EFCRYPSHWRPLEHASRGPNSPYSESYYNSLAVVLQRRDWENPGV
                     TQLNRLAAHPPFASWRNSEEARTDRPSQQLRSL"
     promoter        complement(2208..2226)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2233..2249)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2387..2689
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             3038..3832
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4039..4413
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tgggcttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat ttaggtgaca
     1021 ctatagaata ctcaagctat gcatcaagct tggtaccttg gaccctcgta cagaagctaa
     1081 tacgactcac tatagggaaa taagagagaa aagaagagta agaagaaata taagagccac
     1141 catgagtgtg gatccagctt gtccccaaag cttgccttgc tttgaagcat ccgactgtaa
     1201 agaatcttca cctatgcctg tgatttgtgg gcctgaagaa aactatccat ccttgcaaat
     1261 gtcttctgct gagatgcctc acacggagac tgtctctcct cttccttcct ccatggatct
     1321 gcttattcag gacagccctg attcttccac cagtcccaaa ggcaaacaac ccacttctgc
     1381 agagaagagt gtcgcaaaaa aggaagacaa ggtcccggtc aagaaacaga agaccagaac
     1441 tgtgttctct tccacccagc tgtgtgtact caatgataga tttcagagac agaaatacct
     1501 cagcctccag cagatgcaag aactctccaa catcctgaac ctcagctaca aacaggtgaa
     1561 gacctggttc cagaaccaga gaatgaaatc taagaggtgg cagaaaaaca actggccgaa
     1621 gaatagcaat ggtgtgacgc agaaggcctc agcacctacc taccccagcc tttactcttc
     1681 ctaccaccag ggatgcctgg tgaacccgac tgggaacctt ccaatgtgga gcaaccagac
     1741 ctggaacaat tcaacctgga gcaaccagac ccagaacatc cagtcctgga gcaaccactc
     1801 ctggaacact cagacctggt gcacccaatc ctggaacaat caggcctgga acagtccctt
     1861 ctataactgt ggagaggaat ctctgcagtc ctgcatgcag ttccagccaa attctcctgc
     1921 cagtgacttg gaggctgcct tggaagctgc tggggaaggc cttaatgtaa tacagcagac
     1981 cactaggtat tttagtactc cacaaaccat ggatttattc ctaaactact ccatgaacat
     2041 gcaacctgaa gacgtgtgag ctgccttctg cggggcttgc cttctggcca tgcccttctt
     2101 ctctcccttg cacctgtacc tcttggtctt tgaataaagc ctggaattct gcagatatcc
     2161 atcacactgg cggccgctcg agcatgcatc tagagggccc aattcgccct atagtgagtc
     2221 gtattacaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     2281 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     2341 ccgcaccgat cgcccttccc aacagttgcg cagcctatac gtacggcagt ttaaggttta
     2401 cacctataaa agagagagcc gttatcgtct gtttgtggat gtacagagtg atattattga
     2461 cacgccgggg cgacggatgg tgatccccct ggccagtgca cgtctgctgt cagataaagt
     2521 ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca tgatgaccac
     2581 cgatatggcc agtgtgccgg tctccgttat cggggaagaa gtggctgatc tcagccaccg
     2641 cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataaa tgtcaggcat
     2701 gagattatca aaaaggatct tcacctagat ccttttcacg tagaaagcca gtccgcagaa
     2761 acggtgctga ccccggatga atgtcagcta ctgggctatc tggacaaggg aaaacgcaag
     2821 cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg cgatagctag actgggcggt
     2881 tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggta aggttgggaa
     2941 gccctgcaaa gtaaactgga tggctttctc gccgccaagg atctgatggc gcaggggatc
     3001 aagctctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     3061 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3121 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     3181 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     3241 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     3301 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     3361 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     3421 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     3481 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     3541 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     3601 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     3661 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     3721 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     3781 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gaattattaa
     3841 cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     3901 catacaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     3961 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatag
     4021 cacgtgagga gggccaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc
     4081 gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg
     4141 gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag
     4201 gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg
     4261 tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg
     4321 accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac
     4381 tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc taaaacttca tttttaattt
     4441 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4501 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
//
