LOCUS       Exported                8265 bp ds-DNA     circular SYN 11-JUN-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8265)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jun 11, 2020 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     Ligation of : pSP227_pDZ306_35196.xdna [7443  nt] : (BamHI[2196] / 
            SacI[1690]) to pRS303_Pstl1_qVenus.xdna [814  nt] : (SacI[2073] / 
            BamHI[2891])
FEATURES             Location/Qualifiers
     source          1..8265
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    9..367
                     /label=pGLT1 -550 : -200
     promoter        416..759
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     misc_feature    760..1458
                     /label=S. Klu his3
     terminator      1462..1659
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     misc_feature    1646..1665
                     /label=4242
     misc_feature    1695..2508
                     /label=STL1 promoter
                     /note="note=""Fragment extracted from 
                     pRS303_Pstl1_qVenus.xdna [814  nt] : (SacI[2073] / 
                     BamHI[2891])"
     primer_bind     2277..2296
                     /label=160
     primer_bind     2496..2512
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2509..3838
                     /label=24xPP7 SL
     primer_bind     complement(2603..2619)
                     /label=59
     primer_bind     complement(2724..2740)
                     /label=59
     primer_bind     complement(2845..2861)
                     /label=59
     primer_bind     complement(2966..2982)
                     /label=59
     primer_bind     complement(3087..3103)
                     /label=59
     primer_bind     complement(3208..3224)
                     /label=59
     primer_bind     complement(3329..3345)
                     /label=59
     primer_bind     complement(3450..3466)
                     /label=59
     primer_bind     complement(3571..3587)
                     /label=59
     primer_bind     complement(3692..3708)
                     /label=59
     primer_bind     complement(3813..3829)
                     /label=59
     primer_bind     complement(3934..3950)
                     /label=59
     misc_feature    3999..4303
                     /label=Glt1 +1: 465
     primer_bind     complement(4340..4359)
                     /label=T7
     primer_bind     complement(4341..4359)
                     /label=T7
     misc_feature    4363..4506
                     /label=lacZ_a
                     /note="/label=""lacZ_a"" - 
                     /translation=""MGSLAAVFPGSLDPKVPNCLERSRRYGVAPCRSTLETSRAYGLA
                      - 
                     /GCSIPGFIRSMSQDRILLDRSMPVLKSDNFDPLEEAYEGGTIQNYNDEHHLHKSWANV
                      - 
                     /IPDKRGLYDPDYEHDACGVGFVANKHGEQSHKIVTDARYLLVNMTHRGAVSSDGNGDG
                      - /AGILLGIPRANSRPLNSIRPIVSRITIHWPSFYNVVTGKTLALPNLIALQH"
     primer_bind     complement(4366..4383)
                     /label=M13-fwd
     CDS             4520..4822
                     /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="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             4610..4819
                     /label=ccdB
     promoter        5033..5082
                     /label=NEOKAN_promoter
                     /note="/label=""NEOKAN_promoter"" - 
                     /translation=""MGSLAAVFPGSLDPKVPNCLERSRRYGVAPCRSTLETSRAYGLA
                      - 
                     /GCSIPGFIRSMSQDRILLDRSMPVLKSDNFDPLEEAYEGGTIQNYNDEHHLHKSWANV
                      - 
                     /IPDKRGLYDPDYEHDACGVGFVANKHGEQSHKIVTDARYLLVNMTHRGAVSSDGNGDG
                      - /AGILLGIPRANSRPLNSIRPIVSRITIHWPSFYNVVTGKTLA"
     CDS             5174..5965
                     /label=Kan/neoR
     gene            complement(6215..7073)
                     /label=Ampicillin_Ampicillin
                     /note="/label=""Ampicillin"" - /gene=""Ampicillin"" - 
                     /translation=""MRKEKIPHQEIVSVNNSEELVKKAIEGDALRIGSGDTVKHEEAV
                      - 
                     /SPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILAV
                      - 
                     /GHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQP
                      - /PHCISHDGYFLGRSKVR*"" - 
com"
     rep_origin      7175..7803
                     /direction=RIGHT
                     /label=ColE1 origin
     protein_bind    8073..8094
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8109..8139
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     misc_binding    8143..8165
                     /label=LacO
     primer_bind     8171..8191
                     /label=M13-rev
     promoter        8208..8227
                     /label=T3_promoter
                     /note="/label=""T3_promoter"" - 
                     /translation=""MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                      - 
                     /IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                      - 
                     /YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                      - /DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLI"
ORIGIN
        1 ttaattaaac cgttcaaccc tatacagcgc aaatttggtc ctaatacact tcggttttaa
       61 tgcgtcaatc cgattggctc cgataagctt ttgcacattt ttcaagtcat atgtcacgac
      121 gaacgtgatg cccgcaaaac gtaaaaaaaa gaagcaataa attgcccaaa aaaaaaatat
      181 aaactgcttt tcttagtcat tcataagttc tgatggtctt ccttatctta ttgtattttt
      241 cgtcctattg tttcatttct taccaaatta atcaattctt atatcttact tgataacaca
      301 ccaaactaat cgtctccaca tcataggaag ataggaaatt gctatctcag tcctatacta
      361 cgcagacctc gagtctgttt agcttgcctc gtccccgccg ggtcacccgg ccagcgacat
      421 ggaggcccag aataccctcc ttgacagtct tgacgtgcgc agctcagggg catgatgtga
      481 ctgtcgcccg tacatttagc ccatacatcc ccatgtataa tcatttgcat ccatacattt
      541 tgatggccgc acggcgcgaa gcaaaaatta cggctcctcg ctgcagacct gcgagcaggg
      601 aaacgctccc ctcacagacg cgttgaattg tccccacgcc gcgcccctgt agagaaatat
      661 aaaaggttag gatttgccac tgaggttctt ctttcatata cttcctttta aaatcttgct
      721 aggatacagt tctcacatca catccgaaca taaacaacca tggcagaacc agcccaaaaa
      781 aagcaaaaac aaactgttca ggagcgcaag gcgtttatct cccgtatcac taatgaaact
      841 aaaattcaaa tcgctatttc gctgaatggt ggttatattc aaataaaaga ttcgattctt
      901 cctgcaaaga aggatgacga tgtagcttcc caagctactc agtcacaggt catcgatatt
      961 cacacaggtg ttggcttttt ggatcatatg atccatgcgt tggcaaaaca ctctggttgg
     1021 tctcttattg ttgaatgtat tggtgacctg cacattgacg atcaccatac taccgaagat
     1081 tgcggtatcg cattagggca agcgttcaaa gaagcaatgg gtgctgtccg tggtgtaaaa
     1141 agattcggta ctgggttcgc accattggat gaggcgctat cacgtgccgt agtcgattta
     1201 tctagtagac catttgctgt aatcgacctt ggattgaaga gagagatgat tggtgattta
     1261 tccactgaaa tgattccaca ctttttggaa agtttcgcgg aggcggccag aattactttg
     1321 catgttgatt gtctgagagg tttcaacgat caccacagaa gtgagagtgc gttcaaggct
     1381 ttggctgttg ccataagaga agctatttct agcaatggca ccaatgacgt tccctcaacc
     1441 aaaggtgttt tgatgtgaag tactgacaat aaaaagattc ttgttttcaa gaacttgtca
     1501 tttgtatagt ttttttatat tgtagttgtt ctattttaat caaatgttag cgtgatttat
     1561 attttttttc gcctcgacat catctgccca gatgcgaagt taagtgcgca gaaagtaata
     1621 tcatgcgtca atcgtatgtg aatgctggtc gctatactgc tgtcgattcg atactaacgc
     1681 cgccatccag agctcgcaga acaccactac tgatactcat tgccaaggct aggaggcacc
     1741 atccgtttca ttttctttga ggtaagccaa tcatgaaata gtatacacat ccataacgga
     1801 cgtacggacg aaataagtgc cgttgtccca ctattccacc gcatttggcc catttggctc
     1861 actttgactc aacttgcgtc attttaactg atatgaaggg tccgactttg tcctttttcg
     1921 gccaccgcat accccacggc gatgcctccg ctacctgcat ttgagtaaca tctccgtttc
     1981 gcggggtatt cggcgctacg tcgcctgttc gagcggctct gttcgttgca tgaaactaaa
     2041 ataagcggaa agtgtccagc catccactac gtcagaaaga aataatggtt gtacactgtt
     2101 tctcggctat ataccgtttt tggttggtta atcctcgcca ggtgcagcta ttgcgcttgg
     2161 ctgcttcgcg atagtagtaa tctgagaaag tgcagatccc ggtaagggaa acacttttgg
     2221 ttcacctttg atagggcttt cattggggca ttcgtaacaa aaaggaagta gatagagaaa
     2281 ttgagaaagc ttaagtgaga tgttttagct tcaattttgt ccccttcaac gctgcttggc
     2341 cttagagggt cagaattgca gttcaggagt agtcacactc atagtatata aacaagccct
     2401 ttattgattt tgaataatta ttttgtatac gtgttctagc atacaagtta gaataaataa
     2461 aaaatagaaa aatagaacat agaaagtttt agacctctag aactagtgga tcctaaggta
     2521 cctaattgcc tagaaaggag cagacgatat ggcgtcgctc cctgcaggtc gactctagaa
     2581 accagcagag catatgggct cgctggctgc agtattcccg ggttcattag atcctaaggt
     2641 acctaattgc ctagaaagga gcagacgata tggcgtcgct ccctgcaggt cgactctaga
     2701 aaccagcaga gcatatgggc tcgctggctg cagtattccc gggttcatta gatcctaagg
     2761 tacctaattg cctagaaagg agcagacgat atggcgtcgc tccctgcagg tcgactctag
     2821 aaaccagcag agcatatggg ctcgctggct gcagtattcc cgggttcatt agatcctaag
     2881 gtacctaatt gcctagaaag gagcagacga tatggcgtcg ctccctgcag gtcgactcta
     2941 gaaaccagca gagcatatgg gctcgctggc tgcagtattc ccgggttcat tagatcctaa
     3001 ggtacctaat tgcctagaaa ggagcagacg atatggcgtc gctccctgca ggtcgactct
     3061 agaaaccagc agagcatatg ggctcgctgg ctgcagtatt cccgggttca ttagatccta
     3121 aggtacctaa ttgcctagaa aggagcagac gatatggcgt cgctccctgc aggtcgactc
     3181 tagaaaccag cagagcatat gggctcgctg gctgcagtat tcccgggttc attagatcct
     3241 aaggtaccta attgcctaga aaggagcaga cgatatggcg tcgctccctg caggtcgact
     3301 ctagaaacca gcagagcata tgggctcgct ggctgcagta ttcccgggtt cattagatcc
     3361 taaggtacct aattgcctag aaaggagcag acgatatggc gtcgctccct gcaggtcgac
     3421 tctagaaacc agcagagcat atgggctcgc tggctgcagt attcccgggt tcattagatc
     3481 ctaaggtacc taattgccta gaaaggagca gacgatatgg cgtcgctccc tgcaggtcga
     3541 ctctagaaac cagcagagca tatgggctcg ctggctgcag tattcccggg ttcattagat
     3601 cctaaggtac ctaattgcct agaaaggagc agacgatatg gcgtcgctcc ctgcaggtcg
     3661 actctagaaa ccagcagagc atatgggctc gctggctgca gtattcccgg gttcattaga
     3721 tcctaaggta cctaattgcc tagaaaggag cagacgatat ggcgtcgctc cctgcaggtc
     3781 gactctagaa accagcagag catatgggct cgctggctgc agtattcccg ggttcattag
     3841 atcctaaggt acctaattgc ctagaaagga gcagacgata tggcgtcgct ccctgcaggt
     3901 cgactctaga aaccagcaga gcatatgggc tcgctggctg cagtattccc gggttcatta
     3961 gatccatgtc ccaggataga attttgttag atagatctat gccagtgttg aaatcagaca
     4021 atttcgatcc attggaagaa gcttacgaag gtgggacaat tcaaaactat aacgatgaac
     4081 accatcttca taaatcttgg gcaaatgtga ttccggacaa acgaggactt tacgaccctg
     4141 attatgaaca tgacgcttgt ggtgtcggtt tcgtagcaaa taagcatggt gaacagtctc
     4201 acaagattgt tactgacgct agatatcttt tagtgaatat gacacatcgt ggtgccgtct
     4261 catctgatgg gaacggtgac ggtgccggta ttctgctagg tatcccaagg gcgaattcgc
     4321 ggccgctaaa ttcaattcgc cctatagtga gtcgtattac aattcactgg ccgtcgtttt
     4381 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     4441 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     4501 gcgcagccta tacgtacggc agtttaaggt ttacacctat aaaagagaga gccgttatcg
     4561 tctgtttgtg gatgtacaga gtgatattat tgacacgccg gggcgacgga tggtgatccc
     4621 cctggccagt gcacgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca
     4681 tatcggggat gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtctccgt
     4741 tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa
     4801 cctgatgttc tggggaatat aaatgtcagg catgagatta tcaaaaagga tcttcaccta
     4861 gatccttttc acgtagaaag ccagtccgca gaaacggtgc tgaccccgga tgaatgtcag
     4921 ctactgggct atctggacaa gggaaaacgc aagcgcaaag agaaagcagg tagcttgcag
     4981 tgggcttaca tggcgatagc tagactgggc ggttttatgg acagcaagcg aaccggaatt
     5041 gccagctggg gcgccctctg gtaaggttgg gaagccctgc aaagtaaact ggatggcttt
     5101 cttgccgcca aggatctgat ggcgcagggg atcaagctct gatcaagaga caggatgagg
     5161 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     5221 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     5281 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     5341 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     5401 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     5461 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     5521 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     5581 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     5641 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     5701 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     5761 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     5821 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     5881 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     5941 tcgccttctt gacgagttct tctgaattat taacgcttac aatttcctga tgcggtattt
     6001 tctccttacg catctgtgcg gtatttcaca ccgcatcagg tggcactttt cggggaaatg
     6061 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     6121 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6181 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6241 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6301 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6361 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggc cgagcgcaga
     6421 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6481 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6541 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6601 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6661 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6721 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6781 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6841 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6901 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6961 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7021 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7081 ctttttcaat attattgaag catttatcag ggttattgtc tcatgaccaa aatcccttaa
     7141 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     7201 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     7261 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     7321 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     7381 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     7441 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     7501 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     7561 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     7621 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7681 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7741 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     7801 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     7861 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     7921 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     7981 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     8041 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     8101 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     8161 caatttcaca caggaaacag ctatgaccat gattacgcca agctcagaat taaccctcac
     8221 taaagggact agtcctgcag gtttaaacga attcgccctt atata
//
