LOCUS       pDY0423_EGFP+Kozak_mini 4778 bp ds-DNA     circular     09-MAR-2021
DEFINITION  .
SOURCE      synthetic DNA construct                                            
  ORGANISM  synthetic DNA construct                                            
FEATURES             Location/Qualifiers
     primer_bind     complement(327..344)
                     /label="pBAD Reverse"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     terminator      497..583
                     /label="rrnB T1 terminator"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     terminator      675..702
                     /label="rrnB T2 terminator"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     rep_origin      760..1442
                     /label="ColE1 origin"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     rep_origin      836..1424
                     /label="ori"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     primer_bind     1325..1344
                     /label="pBR322ori-F"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     misc_feature    1413..1436
                     /label="agcctatggaaaaacgccagcaac"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     primer_bind     1578..1595
                     /label="L4440"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     misc_feature    1610..1750
                     /label="bom"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     primer_bind     complement(1676..1695)
                     /label="pRS-marker"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     primer_bind     1836..1858
                     /label="pGEX 3'"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     source          join(2305..4778,1..2304)
                     /label="source"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    complement(2307..2324)
                     /label="I-SceI site"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    complement(2354..2371)
                     /label="I-SceI site"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    complement(2378..2395)
                     /label="I-SceI site"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     misc_feature    complement(2450..2467)
                     /label="I-SceI site"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     misc_feature    complement(2634..2651)
                     /label="I-SceI site"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     misc_feature    complement(2726..2743)
                     /label="I-SceI site"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     misc_feature    complement(2818..2835)
                     /label="I-SceI site"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     misc_feature    complement(2910..2927)
                     /label="I-SceI site"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     CDS             complement(3006..3023)
                     /label="6xHis"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     protein_bind    3044..3077
                     /label="attB"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    3078..3093
                     /label="PD3267"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     misc_feature    3087..3767
                     /label="max GFP"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     CDS             3087..3767
                     /label="Translation 3087-3767"
     misc_feature    3094..3117
                     /label="EGFP ddPCR probe"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    complement(3127..3143)
                     /label="ddPCR reverse"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     misc_feature    3256..3275
                     /label="GCTACGGCTTCTACCACTTC"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    3769..3820
                     /label="AttP seq Bxb1"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     misc_feature    3795..3795
                     /label="this to change to A"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     misc_feature    complement(3796..3817)
                     /label="PD3268"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     misc_feature    3821..3858
                     /label="attP site"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    complement(3946..4742)
                     /label="NeoR"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     CDS             complement(3946..4742)
                     /label="Translation 4742-3946"
     primer_bind     complement(4057..4076)
                     /label="Neo-F"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     primer_bind     4669..4688
                     /label="Neo-R"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
ORIGIN
        1 tgatcccctg cgccatcaga tccttggcgg cgagaaagcc atccagttta ctttgcaggg
       61 cttcccaacc ttaccagagg gcgccccagc tggcaattcc ggttcgcttg ctgtccataa
      121 aaccgcccag tctagctatc gccatgtaag cccactgcaa gctacctgct ttctctttgc
      181 gcttgcgttt tcccttgtcc agatagccca gtagctgaca ttcatccggg gtcagcaccg
      241 tttctgcgga ctggctttct acgtgctcga ggggggccaa acggtctcca gcttggctgt
      301 tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt
      361 ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
      421 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta
      481 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
      541 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
      601 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
      661 gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct
      721 tttgtttatt tttctaaata cattcaaata tgtatccgct catgaccaaa atcccttaac
      781 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
      841 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
      901 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
      961 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     1021 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     1081 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     1141 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     1201 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     1261 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     1321 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     1381 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     1441 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     1501 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     1561 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     1621 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
     1681 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
     1741 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     1801 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     1861 ttcaccgtca tcaccgaaac gcgcgaggca gcagatcaat tcgcgcgcga aggcgaagcg
     1921 gcatgcataa tgtgcctgtc aaatggacga agcagggatt ctgcaaaccc tatgctactc
     1981 cgtcaagccg tcaattgtct gattcgttac caattatgac aacttgacgg ctacatcatt
     2041 cactttttct tcacaaccgg cacggaactc gctcgggctg gccccggtgc attttttaaa
     2101 tacccgcgag aaatagagtt gatcgtcaaa accaacattg cgaccgacgg tggcgatagg
     2161 catccgggtg gtgctcaaaa gcagcttcgc ctggctgata cgttggtcct cgcgccagct
     2221 taagacgcta atccctaact gctggcggaa aagatgtgac agacgcgacg gcgacaagca
     2281 aacatgctgt gcgacgctgg cgatacatta ccctgttatc cctagatgac attaccctgt
     2341 tatcccagat gacattaccc tgttatccct agatgacatt accctgttat ccctagatga
     2401 catttaccct gttatcccta gatgacatta ccctgttatc ccagatgaca ttaccctgtt
     2461 atccctagat acattaccct gttatcccag atgacatacc ctgttatccc tagatgacat
     2521 taccctgtta tcccagatga cattaccctg ttatccctag atacattacc ctgttatccc
     2581 agatgacata ccctgttatc cctagatgac attaccctgt tatcccagat gacattaccc
     2641 tgttatccct agatacatta ccctgttatc ccagatgaca taccctgtta tccctagatg
     2701 acattaccct gttatcccag atgacattac cctgttatcc ctagatacat taccctgtta
     2761 tcccagatga cataccctgt tatccctaga tgacattacc ctgttatccc agatgacatt
     2821 accctgttat ccctagatac attaccctgt tatcccagat gacataccct gttatcccta
     2881 gatgacatta ccctgttatc ccagatgaca ttaccctgtt atccctagat acattaccct
     2941 gttatcccag atgacatacc ctgttatccc tagatgacat taccctgtta tcccagataa
     3001 actcaatgat gatgatgatg atggtcgaga ctcagcggcc gcggtgccag ggcgtgccct
     3061 tgggctcccc gggCGCGGCC GCCACCATGC CCGCCATGAA GATCGAGTGC CGCATCACCG
     3121 GCACCCTGAA CGGCGTGGAG TTCGAGCTGG TGGGCGGCGG AGAGGGCACC CCCGAGCAGG
     3181 GCCGCATGAC CAACAAGATG AAGAGCACCA AAGGCGCCCT GACCTTCAGC CCCTACCTGC
     3241 TGAGCCACGT GATGGGCTAC GGCTTCTACC ACTTCGGCAC CTACCCCAGC GGCTACGAGA
     3301 ACCCCTTCCT GCACGCCATC AACAACGGCG GCTACACCAA CACCCGCATC GAGAAGTACG
     3361 AGGACGGCGG CGTGCTGCAC GTGAGCTTCA GCTACCGCTA CGAGGCCGGC CGCGTGATCG
     3421 GCGACTTCAA GGTGGTGGGC ACCGGCTTCC CCGAGGACAG CGTGATCTTC ACCGACAAGA
     3481 TCATCCGCAG CAACGCCACC GTGGAGCACC TGCACCCCAT GGGCGATAAC GTGCTGGTGG
     3541 GCAGCTTCGC CCGCACCTTC AGCCTGCGCG ACGGCGGCTA CTACAGCTTC GTGGTGGACA
     3601 GCCACATGCA CTTCAAGAGC GCCATCCACC CCAGCATCCT GCAGAACGGG GGCCCCATGT
     3661 TCGCCTTCCG CCGCGTGGAG GAGCTGCACA GCAACACCGA GCTGGGCATC GTGGAGTACC
     3721 AGCACGCCTT CAAGACCCCC ATCGCCTTCG CCAGATCTCG AGCTCGAGGT GGTTTGTCTG
     3781 GTCAACCACC GCGGTCTCAG TGGTGTACGG TACAAACCCA ccccaactgg ggtaaccttt
     3841 gagttctctc agttgggggt aatcagcatc atgatgtggt accacatcat gatgctgatt
     3901 ataagaatgc ggccgccaca ctctagtgga tctcgagtta ataattcaga agaactcgtc
     3961 aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag
     4021 gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat
     4081 gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc
     4141 attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc
     4201 gtcgggcatg ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc
     4261 ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat
     4321 gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg
     4381 cattgcatca gccatgatgg atactttctc ggcaggagca aggtgtagat gacatggaga
     4441 tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg
     4501 agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct
     4561 tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc
     4621 gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag
     4681 ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc
     4741 atgcgaaacg atcctcatcc tgtctcttga tcagagct
//