LOCUS       Exported                4594 bp ds-DNA     circular SYN 25-FEB-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4594)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Feb 25, 2021 from SnapGene Viewer 5.2.4
            https://www.snapgene.com
COMMENT     Insert from 145 with attb sites fragment.ape 13 to 579
            Backbone from pDONR221.ape 2897 to 651
FEATURES             Location/Qualifiers
     source          1..4594
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1..651
                     /organism="pDONR221"
                     /mol_type="other DNA"
                     /label=source:pDONR221
                     /label=source:pDONR221(1)
     source          669..697
                     /organism="pYPQ145"
                     /mol_type="other DNA"
                     /label=source:pYPQ145
     source          join(698..737,2689..2718)
                     /organism="pYPQ145"
                     /mol_type="other DNA"
                     /label=source:pYPQ145
                     /label=source:pYPQ145(1)
     source          738..2688
                     /organism="pDONR221"
                     /mol_type="other DNA"
                     /label=source:pDONR221
     source          2729..4594
                     /organism="pDONR221"
                     /mol_type="other DNA"
                     /label=source:pDONR221
                     /label=source:pDONR221(2)
     terminator      complement(268..295)
                     /label=rrnB_T2_terminator
     terminator      complement(427..470)
                     /label=rrnB_T1_terminator
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    727..730
                     /label=CTAT
     CDS             complement(1133..1438)
                     /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="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(1783..2441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGR"
     gene            complement(1783..2441)
                     /gene="CAT/CamR"
                     /label=CAT/CamR
     promoter        complement(2442..2544)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     misc_feature    2696..2699
                     /label=TGCA
     protein_bind    complement(2719..2818)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2836..2854)
                     /label=T7_promoter
                     /note="/translation=MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQL
                     DITAF 
                     LKTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETF 
                     SSLWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNV 
                     ANMDNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG 
                     A*"
     primer_bind     complement(2859..2875)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2988..3797
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            2988..3797
                     /gene="KanR2"
                     /label=KanR2
                     /note="/translation=MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQL
                     DITAF 
                     LKTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETF 
                     SSLWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNV 
                     ANMDNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG 
                     A*"
     rep_origin      3944..4532
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
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 caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctta tacaaaagtt gccccatggc gttccctcta gataacgcag gatccccaag
      721 tggtggctat cgagaccgtc gaccgacagc cttccaaatg ttcttcgggt gatgctgcca
      781 acttagtcga ccgacagcct tccaaatgtt cttctcaaac ggaatcgtcg tatccagcct
      841 actcgctatt gtcctcaatg ccgtattaaa tcataaaaag aaataagaaa aagaggtgcg
      901 agcctctttt ttgtgtgaca aaataaaaac atctacctat tcatatacgc tagtgtcata
      961 gtcctgaaaa tcatctgcat caagaacaat ttcacaactc ttatactttt ctcttacaag
     1021 tcgttcggct tcatctggat tttcagcctc tatacttact aaacgtgata aagtttctgt
     1081 aatttctact gtatcgacct gcagactggc tgtgtataag ggagcctgac atttatattc
     1141 cccagaacat caggttaatg gcgtttttga tgtcattttc gcggtggctg agatcagcca
     1201 cttcttcccc gataacggag accggcacac tggccatatc ggtggtcatc atgcgccagc
     1261 tttcatcccc gatatgcacc accgggtaaa gttcacggga gactttatct gacagcagac
     1321 gtgcactggc cagggggatc accatccgtc gcccgggcgt gtcaataata tcactctgta
     1381 catccacaaa cagacgataa cggctctctc ttttataggt gtaaacctta aactgcattt
     1441 caccagcccc tgttctcgtc agcaaaagag ccgttcattt caataaaccg ggcgacctca
     1501 gccatccctt cctgattttc cgctttccag cgttcggcac gcagacgacg ggcttcattc
     1561 tgcatggttg tgcttaccag accggagata ttgacatcat atatgccttg agcaactgat
     1621 agctgtcgct gtcaactgtc actgtaatac gctgcttcat agcatacctc tttttgacat
     1681 acttcgggta tacatatcag tatatattct tataccgcaa aaatcagcgc gcaaatacgc
     1741 atactgttat ctggctttta gtaagccgga tccacgcggc gtttacgccc cccctgccac
     1801 tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc atcacaaacg
     1861 gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg
     1921 cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg
     1981 gtgaaactca cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg
     2041 aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc
     2101 cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa
     2161 acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata
     2221 cggaattccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac
     2281 ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg
     2341 ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg
     2401 gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct
     2461 gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag
     2521 ttggaacctc ttacgtgccg atcaacgtct cattttcgcc aaaagttggc ccagggcttc
     2581 ccggtatcaa cagggacacc aggatttatt tattctgcga agtgatcttc cgtcacaggt
     2641 atttattcgg cgcaaagtgc gtcgggtgat gctgccaact tagtcgacgg tctcttgcaa
     2701 ctagtaaggg cgaattcgac ccagctttct tgtacaaagt tggcattata agaaagcatt
     2761 gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc
     2821 atccagctga tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc
     2881 tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaata atatcatcat
     2941 gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc
     3001 aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata
     3061 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc
     3121 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag
     3181 atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt
     3241 ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat
     3301 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt
     3361 tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat
     3421 ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg
     3481 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc
     3541 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg
     3601 acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc
     3661 aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc
     3721 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc
     3781 tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag agcattacgc
     3841 tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt
     3901 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3961 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4021 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4081 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4141 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4201 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4261 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4321 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4381 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4441 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4501 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4561 tcctggcctt ttgctggcct tttgctcaca tgtt
//
