LOCUS       Exported                3866 bp ds-DNA     circular SYN 05-JUL-2018
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    pMuLE_ENTR_TOP-iRFP_L5-L4_full in silico model
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3866)
  AUTHORS   MMCTLab
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, July 5, 2018 from SnapGene 3.1.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3866
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     terminator      268..295
                     /note="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      387..473
                     /gene="Escherichia coli rrnB"
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     537..553
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    570..665
                     /gene="mutant version of attL"
                     /bound_moiety="LR Clonase(TM)"
                     /note="attL5"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    685..807
                     /note="This region does not align with pTA-Luc backbone"
     misc_feature    699..709
                     /note="TCF binding site"
     misc_feature    714..724
                     /note="TCF binding site"
     misc_feature    729..739
                     /note="TCF binding site"
     misc_feature    745..755
                     /note="TCF binding site"
     misc_feature    760..770
                     /note="TCF binding site"
     misc_feature    775..785
                     /note="TCF binding site"
     misc_feature    790..800
                     /note="TCF binding site"
     promoter        840..871
                     /note="minP"
                     /note="minimal TATA-box promoter with low basal activity"
     CDS             933..934
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /note="luciferase"
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation=""
     CDS             945..1895
                     /codon_start=1
                     /product="phytochrome-based near-infrared fluorescent 
                     protein, originally termed iRFP (Filonov et al., 2011)"
                     /note="iRFP713"
                     /note="name was changed to iRFP713 (Shcherbakova and 
                     Verkhusha, 2013)"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGEVIAEDRCAEVESKLGLHYPASTVPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPVHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     protein_bind    complement(1981..2076)
                     /gene="mutant version of attL"
                     /bound_moiety="LR Clonase(TM)"
                     /note="attL4"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2108..2126)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2131..2147)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2132
                     /note="Sequenced as ""N"""
     misc_feature    2137
                     /note="Sequenced as ""N"""
     CDS             2260..3069
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /note="KanR"
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      3216..3804
                     /direction=RIGHT
                     /note="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 tgtatacaaa
      661 agttgggaga cccaagcttg gtaccgagct cttacgcgag atcaaagggg gtaagatcaa
      721 agggggtaag atcaaagggg cgcgagatca aagggggtaa gatcaaaggg ggtaagatca
      781 aagggggtaa gatcaaaggg gcgcgcccgc gtgctagccc gggctcgaga tctagactct
      841 agagggtata taatggaagc tcgaattcca gcttggcatt ccggtactgt tggtaaaaag
      901 cttggcattc cggtactgtt ggtaaagcca ccatcagctg caccatggcg gaaggatccg
      961 tcgccaggca gcctgacctc ttgacctgcg acgatgagcc gatccatatc cccggtgcca
     1021 tccaaccgca tggactgctg ctcgccctcg ccgccgacat gacgatcgtt gccggcagcg
     1081 acaaccttcc cgaactcacc ggactggcga tcggcgccct gatcggccgc tctgcggccg
     1141 atgtcttcga ctcggagacg cacaaccgtc tgacgatcgc cttggccgag cccggggcgg
     1201 ccgtcggagc accgatcact gtcggcttca cgatgcgaaa ggacgcaggc ttcatcggct
     1261 cctggcatcg ccatgatcag ctcatcttcc tcgagctcga gcctccccag cgggacgtcg
     1321 ccgagccgca ggcgttcttc cgccgcacca acagcgccat ccgccgcctg caggccgccg
     1381 aaaccttgga aagcgcctgc gccgccgcgg cgcaagaggt gcggaagatt accggcttcg
     1441 atcgggtgat gatctatcgc ttcgcctccg acttcagcgg cgaagtgatc gcagaggatc
     1501 ggtgcgccga ggtcgagtca aaactaggcc tgcactatcc tgcctcaacc gtgccggcgc
     1561 aggcccgtcg gctctatacc atcaacccgg tacggatcat tcccgatatc aattatcggc
     1621 cggtgccggt caccccagac ctcaatccgg tcaccgggcg gccgattgat cttagcttcg
     1681 ccatcctgcg cagcgtctcg cccgtccatc tggaattcat gcgcaacata ggcatgcacg
     1741 gcacgatgtc gatctcgatt ttgcgcggcg agcgactgtg gggattgatc gtttgccatc
     1801 accgaacgcc gtactacgtc gatctcgatg gccgccaagc ctgcgagcta gtcgcccagg
     1861 ttctggcctg gcagatcggc gtgatggaag agtgagtcac tagtaacggc cgccagtgtg
     1921 ctggaattct gcagatatcc atcacactgg cggccgctcg agcatgcatc tagaggcacc
     1981 caacttttct atacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg
     2041 aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgca gggcggccgc
     2101 gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc
     2161 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
     2221 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     2281 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     2341 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
     2401 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     2461 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     2521 actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta
     2581 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     2641 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
     2701 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     2761 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     2821 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     2881 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     2941 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     3001 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     3061 tttttctaat cagaattggt taattggttg taacactggc agagcattac gctgacttga
     3121 cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga
     3181 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3241 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3301 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3361 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3421 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     3481 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     3541 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     3601 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     3661 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     3721 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     3781 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     3841 ttttgctggc cttttgctca catgtt
//
