LOCUS       Exported                5016 bp ds-DNA     circular SYN 30-MAY-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    SnapGene DNA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5016)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jun 2, 2017 from SnapGene 3.3.4
            http://www.snapgene.com
COMMENT     RFP Gibsoned into SmaI site of pTZ19u
FEATURES             Location/Qualifiers
     source          1..5016
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     240..259
                     /label=T7
     promoter        240..258
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    361..1317
                     /label=apc10
                     /note="abnormal or inviable vegetative cell"
     CDS             join(419..585,657..983,1042..1117)
                     /codon_start=1
                     /label=apc2 CDS
                     /translation="MAQIRQEALKKQKSETQKSTEGFVDIGNLAQWTCSSEKSGFPIRL
                     VRDDNIDTYWQSDGSQPHTIHIKFVKRVSIKYVSMYLQYTLDESYTPSTLRISAGTGFQ
                     DLEIVTTVQVEEPTGWVHVPVGDFGRNGLLDVHLIQIKILANHQSGKDSHVRLIKIYAP
                     EIEQPAIAVDEIPYTSLQFISRNQLR"
     misc_feature    1318..1436
                     /label=rpl301 spacer
     misc_feature    complement(1437..2436)
                     /label=top2 DNA topoisomerase II
     CDS             complement(1529..2436)
                     /codon_start=1
                     /label=top2 CDS
                     /note="viable but with many defects, including abonormal 
                     mitosis, increased plasmid loss, abnormal rDNA spearation. 
                     So this gene will be kept complete."
                     /translation="KTPKELWLHDLDAFEHAWNKVMDDIQREMLEEEQSSRDFVNRTKK
                     KPRGKSTGTRKPRAIAGSSSSTAVKKEASSESKPSTTNRKQQTLLEFAASKEPEKSSDI
                     NIVKTEDNSHGLSVEENRISKSPGLDSSDSGKSRKRSQSVDSEDAGSKKPVKKIAASAS
                     GRGRKTNKPVATTIFSSDDEDDLLPSSLKPSTITSTKASAKNKGKKASSVKKQSPEDDD
                     DDFIIPGSSSTPKASSTNAEPPEDSDSPIRKRPTRRAAATVKTPIYVDPSFDSMDEPSM
                     QDDSFIVDNDEDVDDYDESD"
     primer_bind     complement(2471..2487)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2559..2627
                     /label=LacZ alpha
     rep_origin      2629..3084
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      2646..2952
                     /label=F1 ori
     promoter        3111..3215
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3216..4076
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             join(3414..3931,3933..4073)
                     /label=AmpR
     rep_origin      4171..4853
                     /label=ColE1 origin
     rep_origin      4247..4835
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct
      241 aatacgactc actataggga aagcttgcat gcctgcaggt cgactctaga ggatcggccg
      301 gcccgcggcc gcacagttta atggttgtat agcgtacaca tttgtaaata tagataagga
      361 agccttggga ggacgttatg tcaacatctc acaagtccgc attgtttaca tttagcttat
      421 ggcgcaaatt cgacaagaag cattgaaaaa acaaaagtct gagacgcaaa aatcaacaga
      481 agggttcgtt gatattggaa atttggctca atggacctgc tcgagcgaaa aatccggatt
      541 tcctatacgc ttggtgcggg atgataatat cgatacatat tggcagtaag tgtatttgta
      601 taaatatagt cagggaagaa aatgttttac taactataat tttctttgtc tttaaggtct
      661 gacggctctc aacctcatac catccatatt aaatttgtaa agcgagtatc gattaagtat
      721 gtgagtatgt accttcaata cacccttgat gaatcttata ctcccagtac tttgcgaata
      781 tcagctggga cagggtttca agatttggaa attgttacaa cagttcaagt cgaggaacca
      841 actggttggg tacatgtgcc agttggtgat tttgggcgaa atggcttgtt agatgtgcat
      901 ttaatacaaa tcaaaatact tgcgaaccat caaagtggaa aggatagtca tgttcgtttg
      961 ataaaaatat atgcgccaga aatgtacgta ttttgcatag caaaatcttc attaatataa
     1021 aatgtgctaa cactgatgca gagagcagcc cgcgatagct gtggatgaaa ttccctacac
     1081 aagtttacaa ttcataagta gaaaccaatt acggtaatgc aaaagtcatg tctctggctc
     1141 tctcaaagaa tcgggttgtc atcgttacaa taataattgt tacgttctcg aatttttatg
     1201 gtttttttct tcttttcccc tattatttat gttacaggtt acttcgttta aaaagaaata
     1261 attaatgata ttatgtttga aacttttcat agtaaaaaaa gagtaagtac acagcgacgc
     1321 gtagcatgtt gtaaaacttt agaaaaatac ttgtagaatg agatgcgttg tagtaacata
     1381 ataattgacg atttatttat tttcgcaact tttactatgc tttctaaata tgggtttaat
     1441 tggaactcgt atcctaccat ttacaatgtt catccaattt tttcagattg tactgtaaat
     1501 agcgtttgaa aacaccaaat tttagaagct aatcactctc atcataatcg tctacatcct
     1561 catcgttatc gacgataaaa gaatcatctt gcatgctggg ttcatccatg ctatcaaacg
     1621 agggatcaac gtaaataggt gttttcactg tagccgctgc tcttctggtt ggcctctttc
     1681 taatcggaga atctgaatct tctggtggct ctgcgttagt cgaactagct tttggagttg
     1741 aactactacc tggaataata aaatcatcat cgtcatcttc aggtgattgt ttctttaccg
     1801 agcttgcttt tttcccttta ttcttcgcag aagccttcgt ggatgttatg gtggaaggtt
     1861 tcaaactgct aggcaacaaa tcatcttcat cgtctgaaga aaatatggta gtagcaactg
     1921 gtttattagt ctttcttcct cttccagacg ccgaggctgc tatttttttg acgggttttt
     1981 tactacctgc gtcttcagag tcaacagatt gacttctttt tcttgatttt ccactatcac
     2041 tgctatccaa tcccgggctc ttagatatgc gattttcttc aactgataag ccatgagagt
     2101 tatcctctgt cttgacaatg tttatgtcag atgatttctc aggttctttc gacgctgcga
     2161 actcaagtaa agtttgttgc tttcgatttg ttgtagatgg tttggattcg ctgctagctt
     2221 cttttttaac agcagtactt gaggaggatc cggcaatagc cctgggtttc ctagtaccag
     2281 tggatttacc tcgaggcttc tttttcgttc gatttacaaa atctctagag gattgctctt
     2341 cttctaacat ttctctctga atatcatcca taaccttatt ccaagcatgc tcaaatgcat
     2401 ccaaatcatg aagccacaat tctttaggag ttttttagcg gccgcgggcc ggcctaccga
     2461 gctcgaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     2521 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     2581 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta
     2641 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2701 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2761 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2821 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     2881 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2941 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     3001 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     3061 acaaaatatt aacgtttaca atttcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3121 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3181 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     3241 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     3301 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     3361 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     3421 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     3481 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3541 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3601 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3661 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3721 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     3781 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     3841 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     3901 ttgctgataa atctggagcc ggtgagcgtg gatctcgcgg tatcattgca gcactggggc
     3961 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4021 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     4081 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4141 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4201 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4261 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4321 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4381 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4441 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4501 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4561 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4621 gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4681 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4741 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4801 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4861 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     4921 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     4981 ccgagcgcag cgagtcagtg agcgaggaag cggaag
//
