LOCUS       Exported                6615 bp ds-DNA     circular SYN 27-MAR-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6615)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Mar 29, 2019 from SnapGene Viewer 4.3.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6615
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(236..864)
                     /direction=LEFT
                     /label=ColE1 origin
     CDS             complement(1016..1675)
                     /label=AmpR
     promoter        complement(1915..1943)
                     /label=Amp prom
     rep_origin      2014..2454
                     /direction=RIGHT
                     /label=F1 ori
     CDS             complement(2461..2529)
                     /label=LacZ alpha
     primer_bind     2599..2617
                     /label=M13-fwd
     primer_bind     2627..2646
                     /label=T7
     misc_feature    2681..2750
                     /label=attB
     misc_feature    2778..2877
                     /label=SA
     misc_feature    2878
                     /label=Ph2-5'
     CDS             2879..2917
                     /codon_start=1
                     /label=(GGS)4-BHI linker Phase2
                     /translation="SGGSGGSGGSGGS"
     CDS             2918..2971
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2972..5617
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4"
                     /product="GAL4 transcriptional activator"
                     /label=GAL4
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVSIDSAAHHDNSTIPLDF
                     MPRDALHGFDWSEEDDMSDGLPFLKTDPNNNGFFGDGSLLCILRSIGFKPENYTNSNVN
                     RLPTMITDRYTLASRSTTSRLLQSYLNNFHPYCPIVHSPTLMMLYNNQIEIASKDQWQI
                     LFNCILAIGAWCIEGESTDIDVFYYQNAKSHLTSKVFESGSIILVTALHLLSRYTQWRQ
                     KTNTSYNFHSFSIRMAISLGLNRDLPSSFSDSSILEQRRRIWWSVYSWEIQLSLLYGRS
                     IQLSQNTISFPSSVDDVQRTTTGPTIYHGIIETARLLQVFTKIYELDKTVTAEKSPICA
                     KKCLMICNEIEEVSRQAPKFLQMDISTTALTNLLKEHPWLSFTRFELKWKQLSLIIYVL
                     RDFFTNFTQKKSQLEQDQNDHQSYEVKRCSIMLSDAAQRTVMSVSSYMDNHNVTPYFAW
                     NCSYYLFNAVLVPIKTLLSNSKSNAENNETAQLLQQINTVLMLLKKLATFKIQTCEKYI
                     QVLEEVCAPFLLSQCAIPLPHISYNNSNGSAIKNIVGSATIAQYPTLPEENVNNISVKY
                     VSPGSVGPSPVPLKSGASFSDLVKLLSNRPPSRNSPVTIPRSTPSHRSVTPFLGQQQQL
                     QSLVPLTPSALFGGANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     3'UTR           5618..6132
                     /label=hsp70 3'UTR
     primer_bind     complement(6110..6126)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             6139..6168
                     /codon_start=1
                     /label=(GGS)4 linker Phase2
                     /translation="GGSGGSGGSG"
     misc_feature    6169..6170
                     /label=Ph2-3'
     misc_feature    6171..6291
                     /label=SD
     misc_feature    complement(6319..6388)
                     /label=attB
     primer_bind     complement(6426..6445)
                     /label=T3
     primer_bind     complement(6463..6481)
                     /label=M13-rev
     misc_binding    complement(6489..6511)
                     /label=LacO
     promoter        complement(6516..6545)
                     /label=lac
ORIGIN
        1 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
       61 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
      121 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
      181 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      241 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
      301 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
      361 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
      421 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
      481 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
      541 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
      601 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
      661 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
      721 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
      781 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
      841 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
      901 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
      961 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     1021 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     1081 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     1141 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     1201 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     1261 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     1321 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     1381 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     1441 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     1501 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     1561 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     1621 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     1681 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     1741 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     1801 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     1861 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     1921 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     1981 catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt
     2041 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     2101 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     2161 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     2221 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     2281 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     2341 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     2401 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     2461 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     2521 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     2581 gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat acgactcact
     2641 atagggcgaa ttggagctcg ctagcgatgt aggtcacggt ctcgaagccg cggtgcgggt
     2701 gccagggcgt gcccttgggc tccccgggcg cgtactccac ctcacccatc tggtccatca
     2761 tgatggtcga ctctagaagt cgatccaaca tggcgacttg tcccatcccc ggcatgttta
     2821 aatatactaa ttattcttga actaatttta atcaaccgat ttatctctct tccgcaggtc
     2881 gggaggttcc ggtggaagcg gaggtagcgg cggatccgaa ggacgcggaa gcctgttgac
     2941 gtgcggagat gtggaagaga atccaggacc gatgaagcta ctgtcttcta tcgaacaagc
     3001 atgcgatatt tgccgactta aaaagctcaa gtgctccaaa gaaaaaccga agtgcgccaa
     3061 gtgtctgaag aacaactggg agtgtcgcta ctctcccaaa accaaaaggt ctccgctgac
     3121 tagggcacat ctgacagaag tggaatcaag gctagaaaga ctggaacagc tatttctact
     3181 gatttttcct cgagaagacc ttgacatgat tttgaaaatg gattctttac aggatataaa
     3241 agcattgtta acaggattat ttgtacaaga taatgtgaat aaagatgccg tcacagatag
     3301 attggcttca gtggagactg atatgcctct aacattgaga cagcatagaa taagtgcgac
     3361 atcatcatcg gaagagagta gtaacaaagg tcaaagacag ttgactgtat cgattgactc
     3421 ggcagctcat catgataact ccacaattcc gttggatttt atgcccaggg atgctcttca
     3481 tggatttgat tggtctgaag aggatgacat gtcggatggc ttgcccttcc tgaaaacgga
     3541 ccccaacaat aatgggttct ttggcgacgg ttctctctta tgtattcttc gatctattgg
     3601 ctttaaaccg gaaaattaca cgaactctaa cgttaacagg ctcccgacca tgattacgga
     3661 tagatacacg ttggcttcta gatccacaac atcccgttta cttcaaagtt atctcaataa
     3721 ttttcacccc tactgcccta tcgtgcactc accgacgcta atgatgttgt ataataacca
     3781 gattgaaatc gcgtcgaagg atcaatggca aatccttttt aactgcatat tagccattgg
     3841 agcctggtgt atagaggggg aatctactga tatagatgtt ttttactatc aaaatgctaa
     3901 atctcatttg acgagcaagg tcttcgagtc aggttccata attttggtga cagccctaca
     3961 tcttctgtcg cgatatacac agtggaggca gaaaacaaat actagctata attttcacag
     4021 cttttccata agaatggcca tatcattggg cttgaatagg gacctcccct cgtccttcag
     4081 tgatagcagc attctggaac aaagacgccg aatttggtgg tctgtctact cttgggagat
     4141 ccaattgtcc ctgctttatg gtcgatccat ccagctttct cagaatacaa tctccttccc
     4201 ttcttctgtc gacgatgtgc agcgtaccac aacaggtccc accatatatc atggcatcat
     4261 tgaaacagca aggctcttac aagttttcac aaaaatctat gaactagaca aaacagtaac
     4321 tgcagaaaaa agtcctatat gtgcaaaaaa atgcttgatg atttgtaatg agattgagga
     4381 ggtttcgaga caggcaccaa agtttttaca aatggatatt tccaccaccg ctctaaccaa
     4441 tttgttgaag gaacaccctt ggctatcctt tacaagattc gaactgaagt ggaaacagtt
     4501 gtctcttatc atttatgtat taagagattt tttcactaat tttacccaga aaaagtcaca
     4561 actagaacag gatcaaaatg atcatcaaag ttatgaagtt aaacgatgct ccatcatgtt
     4621 aagcgatgca gcacaaagaa ctgttatgtc tgtaagtagc tatatggaca atcataatgt
     4681 caccccatat tttgcctgga attgttctta ttacttgttc aatgcagtcc tagtacccat
     4741 aaagactcta ctctcaaact caaaatcgaa tgctgagaat aacgagaccg cacaattatt
     4801 acaacaaatt aacactgttc tgatgctatt aaaaaaactg gccactttta aaatccagac
     4861 ttgtgaaaaa tacattcaag tactggaaga ggtatgtgcg ccgtttctgt tatcacagtg
     4921 tgcaatccca ttaccgcata tcagttataa caatagtaat ggtagcgcca ttaaaaatat
     4981 tgtcggttct gcaactatcg cccaataccc tactcttccg gaggaaaatg tcaacaatat
     5041 cagtgttaaa tatgtttctc ctggctcagt agggccttca cctgtgccat tgaaatcagg
     5101 agcaagtttc agtgatctag tcaagctgtt atctaaccgt ccaccctctc gtaactctcc
     5161 agtgacaata ccaagaagca caccttcgca tcgctcagtc acgccttttc tagggcaaca
     5221 gcaacagctg caatcattag tgccactgac cccgtctgct ttgtttggtg gcgccaattt
     5281 taatcaaagt gggaatattg ctgatagctc attgtccttc actttcacta acagtagcaa
     5341 cggtccgaac ctcataacaa ctcaaacaaa ttctcaagcg ctttcacaac caattgcctc
     5401 ctctaacgtt catgataact tcatgaataa tgaaatcacg gctagtaaaa ttgatgatgg
     5461 taataattca aaaccactgt cacctggttg gacggaccaa actgcgtata acgcgtttgg
     5521 aatcactaca gggatgttta ataccactac aatggatgat gtatataact atctattcga
     5581 tgatgaagat accccaccaa acccaaaaaa agagtaaaat gaatcgtaga tactgaaaaa
     5641 ccccgcaagt tcacttcaac tgtgcatcgt gcaccatctc aatttctttc atttatacat
     5701 cgttttgcct tcttttatgt aactatactc ctctaagttt caatcttggc catgtaacct
     5761 ctgatctata gaatttttta aatgactaga attaatgccc atcttttttt tggacctaaa
     5821 ttcttcatga aaatatatta cgagggctta ttcagaagct tatcgatacc gtcgactaaa
     5881 gccaaataga aattattcag ttctggctta agtttttaaa agtgatatta tttatttggt
     5941 tgtaaccaac caaaagaatg taaataacta atacataatt atgttagttt taagttagca
     6001 acaaattgat tttagctata ttagctactt ggttaataaa tagaatatat ttatttaaag
     6061 ataattcgtt tttattgtca gggagtgagt ttgcttaaaa actcgtttag atccactagt
     6121 tctagagcgg ccggatccgg aggtagcggt ggaagcggag gttccggcag gtaagttatt
     6181 gaacaatggc atcaaatgcc ttcatcatca ctacccttta gcccttaaga ccccacaatg
     6241 accttaccca ctcagagaaa aaagtaaata tgaaagccca tttgaacttc taagcttgtc
     6301 gaccatcatg atggaccaga tgggtgaggt ggagtacgcg cccggggagc ccaagggcac
     6361 gccctggcac ccgcaccgcg gcttcgagac cgtgacctac atcatgcatg gtacccagct
     6421 tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc
     6481 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6541 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6601 ccgctttcca gtcgg
//
