LOCUS       Exported                4443 bp ds-DNA     circular SYN 25-MAR-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4443)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Mar 25, 2019 from SnapGene Viewer 4.3.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4443
                     /organism="synthetic DNA construct"
                     /note="/dnas_title=pDONR221_Trf2_noATG-STOP_Anderson"
     regulatory      complement(258..295)
                     /regulatory_class="terminator"
                     /note="rrnB transcription terminator T2"
                     /note="transcription terminator for E. coli RNA polymerase"
                     /note="/dnas_title=rrnB T2 terminator"
     regulatory      complement(421..477)
                     /regulatory_class="terminator"
                     /note="rrnB transcription terminator T1"
                     /note="transcription terminator for E. coli RNA polymerase"
                     /note="/dnas_title=rrnB T1 terminator"
     misc_recomb     570..668
                     /label=attL1
                     /note="attL1"
                     /note="recombination site for the Gateway(R) BP reaction"
                     /note="recognized as attL1"
                     /note="/dnas_title=attL1"
     misc_feature    659..2577
                     /label=nonstandard type: PCR_product
                     /note="/current=1"
                     /note="/pair=AttB PCR product"
                     /note="/score=1.000"
                     /note="/set=Set 1"
     misc_feature    659..2577
                     /label=nonstandard type: clone
                     /note="/source=pDONR223-Trf2(671,2563) AttB Insert"
                     /note="/type="
     primer_bind     659..689
                     /label=Primer 1
     misc_feature    671..2566
                     /label=Trf2-short
                     /note="Trf2-short"
     misc_feature    671..2563
                     /label=nonstandard type: insert
                     /note="/source=pDONR223-Trf2(671,2563)"
                     /note="/type=BP AttB Insert"
     misc_feature    671..2563
                     /label=nonstandard type: PCR_frame
                     /note="Reading Frame"
                     /note="/set=Set 1"
     primer_bind     complement(2541..2577)
                     /label=Primer 2
     misc_recomb     complement(2568..2666)
                     /label=attL2
                     /note="attL2"
                     /note="recombination site for the Gateway(R) BP reaction"
                     /note="recognized as attL2"
                     /note="/dnas_title=attL2"
     misc_feature    join(2578..4443,1..658)
                     /label=nonstandard type: vector
                     /note="/source=pDONR221"
                     /note="/type=Donor Vector"
     regulatory      complement(2686..2703)
                     /regulatory_class="promoter"
                     /note="T7 promoter"
                     /note="bacteriophage T7 RNA polymerase promoter"
                     /note="/dnas_title=T7 promoter"
     CDS             2837..3646
                     /codon_start=1
                     /gene="nptI"
                     /gene_synonym="kan; neo; aphA1; aph(3')-Ia"
                     /product="Aminoglycoside 3'-phosphotransferase"
                     /function="confers resistance to neomycin, kanamycin, and 
                     G418 (Geneticin(R))"
                     /EC_number="2.7.1.95"
                     /label=nptI
                     /note="APH(3')-I; Kanamycin kinase, type I; 
                     Neomycin-kanamycin phosphotransferase type I; APH(3')I"
                     /note="/dnas_title=KanR"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      3770..4391
                     /label=pMB1 origin of replication
                     /note="pMB1 origin of replication"
                     /note="C171T is a pUC mutation; none of the other observed 
                     variants are described in the literature"
                     /note="/dnas_title=pMB1 ori"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggcttc caaaacgata tggtcagcat accggtggcc aatttgaacg gcggccttaa
      721 agcggccagc agcggttccg gagttggtgt agtgacctcc ggcggcgtcg tatcctcggc
      781 ggtgctggcc aatgccccac gtgtatacct aacgccctcc tccacattca tgaccaatcg
      841 ccaaatggcg ggagtggcgt ccaccggaag aatgagcgga caagtcgtcg gtggttcatc
      901 tggcacagcc tcaacagccg gtaccgttcg ctatttttcg caattcagca aaatgcaaac
      961 ggccggcgga cccagtttgc aaaggaaact tgccaacggg gataccattg ttctggcaac
     1021 cggcagtaaa aatatgttcc ttactagcag cgaaaataag gctaatctgc cgaccgtcgc
     1081 cagcaatggc aatggtctga tcacagccaa aatggatcta ttggaggagg aggtcatgca
     1141 gtcgataact gtgattgatg acgatgacga ggagaaaaag gaagtcgctg aagacgaaga
     1201 agagagcagc aataatgcca aacccattga ccttcaccag ccaattgctg ataatgagca
     1261 cgagctggac attgtgataa acaatgtggt ctgctcgttt agcgtcggct gtcacctgaa
     1321 gctgcgcgaa attgcgctgc agggatcaaa tgtggagtac cgtcgcgaga acggaatggt
     1381 gacaatgaag ctccgccatc catacaccac tgcctcgatt tggtcgtcgg ggaggattac
     1441 ctgcactggt gccacttccg agtcaatggc caaggtcgct gcacgccgct atgccagatg
     1501 ccttggcaag ctgggatttc ccactcgttt cctcaacttt cgcattgtca acgtactggg
     1561 cacctgcagc atgccgtggg ccatcaagat cgtcaacttt tcggagcgcc atcgcgagaa
     1621 cgccagctac gagcccgagc tgcatcctgg agtgacttac aagatgcgcg atcccgatcc
     1681 aaaggccacc ctgaagatct tctccaccgg cagcgtaaca gtcaccgcgg ctagcgtaaa
     1741 ccacgttgaa tcggctatcc agcacatata cccgctggtc tttgatttcc gcaagcaacg
     1801 ttcggcggag gagctgcagc atctgcgtca aaagcagcgt ctgcaagccg gcggtgatcc
     1861 ccacgaactg gagaaaaatg tcttggccga taacaagact gcttcccttg ataatatctt
     1921 tgttaataca acagcggcgc actctaaatc atcgtcgaat gaccaaacat cagcacccgc
     1981 gaccatcttg tccagcacag ttgatagtat gcctcgtctg aaacagatgg tcaactatca
     2041 ccagatgatg aagcaaaccc aggaagagcg ccgccatata atgtttaatg gggagaaggc
     2101 caatccggcc tccacctcat cggcggcagc agctccctcc acctcctcct catcgtcctc
     2161 atccggggat aatatctgtg ccaatgcccg tcgccgggcc accgaatgct gggcaaccaa
     2221 gctgcagaat aagcgtccac gttacaatga tcccggtacc acgggcacaa ttaacgccgc
     2281 atcgtccaca gcatcagccg ccacatcttc gttagcctcc caggcgacac atctgcgtaa
     2341 tcctctcaag acggccgcgt tggccaatgc acgtatgtta ggcgccaagg tgacaacatg
     2401 cacacgcaac agcataatcg tgcagcagcc gcaacgtatc caaatgcagc agcagcagca
     2461 acagctgcaa ccgcagcagc aacagaccag cttctcgccc agcgaattcg atgttgatga
     2521 cttgatcgag gaggaggaga acaacgaatt ggatatgccc ttctaggacc cagctttctt
     2581 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     2641 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     2701 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     2761 cattgcacaa gataaaataa tatcatcatg aacaataaaa ctgtctgctt acataaacag
     2821 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     2881 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     2941 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     3001 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     3061 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     3121 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     3181 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     3241 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     3301 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     3361 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     3421 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     3481 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     3541 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     3601 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     3661 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     3721 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     3781 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3841 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3901 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3961 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4021 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4081 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4141 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4201 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4261 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4321 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4381 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4441 gtt
//
