LOCUS       Exported                5714 bp ds-DNA     circular SYN 21-NOV-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5714)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 21, 2020 from SnapGene 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5714
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          178..280
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label=source:synthetic DNA construct
     source          281..583
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label=source:synthetic DNA construct
                     /note="ApEinfo_fwdcolor=pink
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_revcolor=pink"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     RBS             59..71
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    81..166
                     /label=MalE signal sequenced optimized
     promoter        178..280
                     /label=PccdB
                     /label=ccdB insert
     CDS             281..583
                     /codon_start=1
                     /label=ccdB
                     /note="ccdB
                     ApEinfo_fwdcolor=#f63cff
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} 
                     width 5 offset 0
                     ApEinfo_revcolor=#00ff00"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDKSWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     terminator      659..706
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      743..1198
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(1291..2106)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      2228..2816
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3002..3144
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3246..3437)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4246..5328)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(5329..5406)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atcgtattat gaaaattaaa acaggtgcac gcatcctcgc
      121 attatccgca ttaacgacga tgatgttttc cgcctcggct ctcgcccatc gaagagctac
      181 taaaagccag ataacagtat gcgtatttgc gcgctgattt ttgcggtata agaatatata
      241 ctgatatgta tacccgaagt atgtcaaaaa gaggtatgct atgcagttta aggtttacac
      301 ctataaaaga gagagccgtt atcgtctgtt tgtggatgta cagagtgata ttattgacac
      361 gcccgggcga cggatggtga tccccctggc cagtgcacgt ctgctgtcag ataaagtctc
      421 ccgtgaactt tacccggtgg tgcatatcgg ggataaaagc tggcgcatga tgaccaccga
      481 tatggccagt gtgccggtct ccgttatcgg ggaagaagtg gctgatctca gccaccgcga
      541 aaatgacatc aaaaacgcca ttaacctgat gttctgggga ataggctctt cgtaagatcc
      601 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
      661 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
      721 tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
      781 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
      841 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
      901 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
      961 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     1021 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     1081 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     1141 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttca
     1201 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     1261 tcaaatatgt atccgctcat gaattaattc ttagaaaaac tcatcgagca tcaaatgaaa
     1321 ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa
     1381 tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc
     1441 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt
     1501 atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg
     1561 catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc
     1621 atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct
     1681 gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc
     1741 atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc
     1801 ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt
     1861 cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt
     1921 ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa
     1981 tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa
     2041 atcagcatcc atgttggaat ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg
     2101 gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgacca
     2161 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     2221 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     2281 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     2341 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     2401 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     2461 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     2521 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     2581 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     2641 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     2701 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     2761 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     2821 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     2881 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     2941 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagg
     3001 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatatggtg
     3061 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     3121 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     3181 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     3241 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
     3301 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg
     3361 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
     3421 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
     3481 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
     3541 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
     3601 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
     3661 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
     3721 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
     3781 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca
     3841 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc
     3901 cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga
     3961 ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg
     4021 atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc
     4081 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg
     4141 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
     4201 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
     4261 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4321 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     4381 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     4441 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     4501 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
     4561 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     4621 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     4681 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     4741 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     4801 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     4861 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     4921 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     4981 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     5041 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     5101 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     5161 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     5221 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     5281 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     5341 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     5401 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta
     5461 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
     5521 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     5581 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     5641 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct
     5701 cgatcccgcg aaat
//
