LOCUS       40924_21245        6680 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pGCD21, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6680)
  AUTHORS   Dantas GC, Martins PMM., Ferreira H.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-FEB-2016) Bioquimica e Microbiologia, UNESP, Av. 24A, 
            1515, Rio Claro, SP 13506900, Brazil
REFERENCE   2  (bases 1 to 6680)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6680)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (04-FEB-2016) Bioquimica e Microbiologia, UNESP, Av. 24A, 1515, Rio 
            Claro, SP 13506900, Brazil"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6680
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(98..973)
                     /label=araC
                     /note="L-arabinose regulatory protein"
     promoter        1000..1284
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     regulatory      1305..1310
                     /regulatory_class="ribosome_binding_site"
     CDS             1331..2044
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
     misc_feature    complement(2081..2895)
                     /label=Xac_amy
                     /note="Xac_amy"
     primer_bind     complement(2908..2924)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3065..3493
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             3837..4628
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             4649..5506
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      5680..6268
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6556..6577
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6592..6622
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6630..6646
                     /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     6654..6670
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aattcgcata atgtgcctgt caaatggacg aagcagggat tctgcaaacc ctatgctact
       61 ccgtcaagcc gtcaattgtc tgattcgtta ccaattatga caacttgacg gctacatcat
      121 tcactttttc ttcacaaccg gcacggaact cgctcgggct ggccccggtg cattttttaa
      181 atacccgcga gaaatagagt tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag
      241 gcatccgggt ggtgctcaaa agcagcttcg cctggctgat acgttggtcc tcgcgccagc
      301 ttaagacgct aatccctaac tgctggcgga aaagatgtga cagacgcgac ggcgacaagc
      361 aaacatgctg tgcgacgctg gcgatatcaa aattgctgtc tgccaggtga tcgctgatgt
      421 actgacaagc ctcgcgtacc cgattatcca tcggtggatg gagcgactcg ttaatcgctt
      481 ccatgcgccg cagtaacaat tgctcaagca gatttatcgc cagcagctcc gaatagcgcc
      541 cttccccttg cccggcgtta atgatttgcc caaacaggtc gctgaaatgc ggctggtgcg
      601 cttcatccgg gcgaaagaac cccgtattgg caaatattga cggccagtta agccattcat
      661 gccagtaggc gcgcggacga aagtaaaccc actggtgata ccattcgcga gcctccggat
      721 gacgaccgta gtgatgaatc tctcctggcg ggaacagcaa aatatcaccc ggtcggcaaa
      781 caaattctcg tccctgattt ttcaccaccc cctgaccgcg aatggtgaga ttgagaatat
      841 aacctttcat tcccagcggt cggtcgataa aaaaatcgag ataaccgttg gcctcaatcg
      901 gcgttaaacc cgccaccaga tgggcattaa acgagtatcc cggcagcagg ggatcatttt
      961 gcgcttcagc catacttttc atactcccgc cattcagaga agaaaccaat tgtccatatt
     1021 gcatcagaca ttgccgtcac tgcgtctttt actggctctt ctcgctaacc aaaccggtaa
     1081 ccccgcttat taaaagcatt ctgtaacaaa gcgggaccaa agccatgaca aaaacgcgta
     1141 acaaaagtgt ctataatcac ggcagaaaag tccacattga ttatttgcac ggcgtcacac
     1201 tttgctatgc catagcattt ttatccataa gattagcgga tcctacctga cgctttttat
     1261 cgcaactctc tactgtttct ccatacccgt ttttttgggc tagcaggagg aaagatccat
     1321 ggcactcgag atgagtaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga
     1381 attagatggt gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc
     1441 aacatacgga aaacttaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg
     1501 gccaacactt gtcactactt tcgcgtatgg tcttcaatgc tttgcgagat acccagatca
     1561 tatgaaacag catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaagaac
     1621 tatatttttc aaagatgacg ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga
     1681 tacccttgtt aatagaatcg agttaaaagg tattgatttt aaagaagatg gaaacattct
     1741 tggacacaaa ttggaataca actataactc acacaatgta tacatcatgg cagacaaaca
     1801 aaagaatgga atcaaagtta acttcaaaat tagacacaac attgaagatg gaagcgttca
     1861 actagcagac cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga
     1921 caaccattac ctgtccacac aatctgccct ttcgaaagat cccaacgaaa agagagacca
     1981 catggtcctt cttgagtttg taacagctgc tgggattaca catggcatgg atgaactata
     2041 caaagaagca ggcggccgcg cactgaatct agaatgaccc gggagctgct gcatgctggc
     2101 gccaatggcg aacgcgttgc gcaccgcatt gaacagcgga aagtcgtagg ccgcatgcgg
     2161 cgtggattgc aggtacggcg ccaggaactg gtcgtaatcg ccattgcccg taccgccgcc
     2221 ggtgatcact tcgccgaaga catacacacc cgatcggatg ccggcatcga agacgcgatt
     2281 gagatggtcg aacgtcatgt gcttggccgc atccacccgg aagccggtca cgcccatgct
     2341 cttgagcgcc tgcagatagg cgcgctgctg ctgtaccacc cagtcgttgc ccagcaaatc
     2401 cggcaggcct ggatcgctgc cgccgccgca gatacggtaa ttgcgcacct ggtaagcgtc
     2461 gttgtagttg ctgatgcact gcgccgggcc gaaatcgctg gcgctgagga agtttgactg
     2521 cagcgtgccg aacaagcgca gcgaatcgta acggccgggg tttgccgcgt actgcgacag
     2581 gaccgcgccg cctgggtaat tgaggtccga gcgtgtggcc gcttcgttgg ccatgtggtt
     2641 gaacacgatg tcggcatagg tttccacgcc gttgctggcc agcgcctgga ccatcttctt
     2701 gaatgcagcg gtgtcgccga gcggattgtc gatcaggcgg atgtcctggg gttgatagcg
     2761 cgcccaccag gcacttcctt cggagcggta cgcgggggcg accaatacct tgcgatagcc
     2821 ggcatcggca atctgcttgg cgcgcgcttc cacggtggca tacggccagt tgaaggcgtg
     2881 caggatgaca tccccgggaa gcttggcact ggccgtcgtt ttacaacgtc gtgactggga
     2941 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     3001 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     3061 atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3121 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3181 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3241 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3301 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3361 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3421 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3481 tttaacgcga attttaacaa aattcagggc gcaagggctg ctaaaggaag cggaacacgt
     3541 agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac tgggctatct
     3601 ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg cttacatggc
     3661 gatagctaga ctgggcggtt ttatggacag caagcgaacc ggaattgcca gctggggcgc
     3721 cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga
     3781 tctgatggcg caggggatca agatctgatc aagagacagg atgaggatcg tttcgcatga
     3841 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3901 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3961 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     4021 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4081 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4141 tcctgtcatc ccaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4201 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4261 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4321 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     4381 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4441 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4501 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4561 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4621 agttcttctg aattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4681 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4741 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4801 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4861 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     4921 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4981 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5041 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5101 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5161 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     5221 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5281 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5341 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5401 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5461 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5521 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5581 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5641 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5701 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5761 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5821 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5881 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5941 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6001 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6061 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6121 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6181 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6241 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6301 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6361 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     6421 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     6481 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     6541 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     6601 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     6661 cagctatgac catgattacg
//
