LOCUS       Exported                4965 bp ds-DNA     circular SYN 06-JUN-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4965)
  AUTHORS   Weatherall Institute
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Mar 22, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4965
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        255..503
                     /label=Human U6 Promoter
     misc_feature    483..515
                     /label=Feature 22
     primer_bind     499..508
                     /label=TM1-Emp-amp-F_NEW
     misc_feature    500..503
                     /label=RE-BbsI
     misc_feature    504..515
                     /label=4nt hairpin
     primer_bind     516..560
                     /label=X-T-Y-1F
     misc_feature    516..519
                     /label=bbsI
     misc_feature    516..517
                     /label=4nt hairpin
     misc_feature    518..519
                     /label=link
     primer_bind     complement(520..566)
                     /label=X-T-Y-1*R
     misc_feature    521..526
                     /label=BsmBI
     misc_feature    527..542
                     /label=random
     misc_feature    532..537
                     /label=XhoI
     misc_feature    543..548
                     /label=BsmBI
     misc_feature    550..553
                     /label=new-bbsI
     misc_feature    554..599
                     /label=trigger place holder
     primer_bind     561..607
                     /label=X-T-Y-2F
     misc_feature    562..591
                     /label=random
                     /note="added to facilitate cloning (longer product)"
     primer_bind     complement(567..611)
                     /label=X-T-Y-2*R
     misc_feature    574..579
                     /label=XbaI
     misc_feature    600..607
                     /label=NotI
     misc_feature    600..601
                     /label=!!!Needs to be there
     misc_feature    608..611
                     /label=bbsI
     misc_feature    608..609
                     /label=link
     misc_feature    610..611
                     /label=8nt hairpin
     misc_feature    612..648
                     /label=Feature 23
     misc_feature    612..629
                     /label=8nt hairpin
     terminator      630..635
                     /label=U6-TER
                     /note="U6 terminator"
     promoter        1196..1505
                     /label=SV40 promoter
                     /note="SV40 promoter"
     primer_bind     1348..1366
                     /label=ibIns_bk_1_F_62
     misc_feature    1381..1386
                     /label=Feature 17
     primer_bind     1506..1546
                     /label=ibIns_i_3_F_63
     misc_feature    1506..1529
                     /label=Feature 18
     primer_bind     complement(1506..1529)
                     /label=ibIns_bk_2_R_61
     promoter        1506..1520
                     /label=SV40 promoter
                     /note="SV40 promoter"
     primer_bind     1530..1546
                     /label=ibIns_i_3_F_*
     misc_feature    1530..1538
                     /label=KS
     CDS             1536..2498
                     /codon_start=1
                     /label=iBlue
                     /translation="MSVPLTTSAFGHAFLANCEREQIHLAGSIQPHGILLAVKEPDNVV
                     IQASINAAEFLNTNSVVGRPLRDLGGDLPLQILPHLNGPLHLAPMTLRCTVGSPPRRVD
                     CTIHRPSNGGLIVELEPATKTTNIAPALDGAFHRITSSSSLMGLCDETATIIREITGYD
                     RVMVVRFDEEGNGQILSERRRADLEAFLGNRYPASTIPQIARRLYEHNRVRLLVDVNYT
                     PVPLQPRISPLNGRDLDMSLSCLRSMSPCHQKYMQDMGVGATLVCSLMVSGRLWGLIAC
                     HHYEPRFVPFHIRAAGEALAETCAIRIATLESFAQSQSK"
     primer_bind     complement(2478..2504)
                     /label=ibIns_i_4_R_63
     primer_bind     complement(2478..2498)
                     /label=ibIns_i_4_R_63*
     misc_feature    2499..2504
                     /label=Feature 16
     CDS             2500..2509
                     /codon_start=1
                     /label=ECFP
                     /translation="YK"
     polyA_signal    2522..2894
                     /label=SV40pA
                     /note="SV40pA"
     primer_bind     complement(2789..2810)
                     /label=Primer 6
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagtgagggc ctatttccca tgattccttc atatttgcat atacgataca
      301 aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata ttagtacaaa
      361 atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta
      421 aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata
      481 tcttgtggaa aggacgaaac accggctcga aagagccact gagacgtaag actcgagcgg
      541 tccgtctcag tcagggtctt cgagcattgg aagtctagat acctgcttta cgaagacctg
      601 cggccgcatg ctagtacgaa agtactagct tttttgctag caggcatgct ggggatgcgg
      661 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
      721 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
      781 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
      841 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
      901 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
      961 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     1021 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     1081 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     1141 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     1201 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     1261 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     1321 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     1381 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     1441 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     1501 agcttgtata tccattttcg gatctgatcg ccaccatgtc ggtaccgctg actacctcag
     1561 cattcggcca cgcgtttctg gctaactgtg aacgcgagca gatccacctg gcgggctcca
     1621 ttcagccgca cggtatcctg ctggctgtga aagagccgga caacgtggtg atccaggctt
     1681 ctattaacgc tgcggagttc ctgaacacca actctgttgt tggccgtccg ctgcgtgacc
     1741 tgggcggcga tctgcctttg cagatcctgc cgcacctgaa cggcccgctg cacctggctc
     1801 cgatgaccct gcgttgtacc gtgggttctc cgccgcgtcg tgtggactgt accattcatc
     1861 gtccgtctaa cggcggcctg atcgtagaac tggaaccagc aaccaagacc actaacattg
     1921 cgccggctct ggacggtgcg tttcatcgta tcacttcttc atcctccctg atgggcctgt
     1981 gtgacgaaac cgcgactatt atccgtgaga ttactggcta cgaccgtgtg atggtagtac
     2041 gtttcgatga agagggtaat ggccaaattc tgtccgaacg tcgtcgtgcg gacctggaag
     2101 cgttcctggg taaccgctac ccggcgtcta ctattccgca gatcgctcgt cgcctgtacg
     2161 aacataaccg tgttcgcctg ctggtagatg tgaactatac tccggttccg ctacagccgc
     2221 gcatcagccc gctgaacggt cgtgatctgg atatgtccct gtcttgcctg cgctctatgt
     2281 ccccgtgcca ccagaaatac atgcaggaca tgggcgttgg cgcgaccctg gtttgctctc
     2341 tgatggtgtc tggtcgtctg tggggtctga tcgcttgcca ccactacgaa ccgcgcttcg
     2401 ttccgttcca cattcgcgct gctggcgaag cgctggcgga aacttgtgcg atccgcatcg
     2461 cgacgctgga gagctttgca cagtctcagt ccaaatgatg tacaagtaaa gcacgtgcta
     2521 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     2581 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     2641 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     2701 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     2761 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     2821 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     2881 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     2941 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     3001 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     3061 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3121 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3181 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3241 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3301 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3361 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3421 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3481 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3541 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3601 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3661 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3721 tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc
     3781 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3841 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3901 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3961 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4021 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4081 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4141 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4201 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4261 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4321 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4381 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4441 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4501 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4561 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4621 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4681 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4741 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4801 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     4861 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4921 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
