LOCUS       pET22b-mEB1-eGFP-6xHis_MBN.xdna    6909 bp      DNA     circular   19/10/2019
DEFINITION  
ACCESSION   
VERSION     
SOURCE      
  ORGANISM  
COMMENT     Serial Cloner Genbank Format
COMMENT     SerialCloner_Type=DNA
COMMENT     SerialCloner_Comments=[DEFINITION]: synthetic circular DNA - [LOCUS]: Exported - [
            ACCESSION]: . - [GENE]: complement(899..1702) - [CDS]: compl
            ement(140..157)
COMMENT     SerialCloner_Ends=0,0,,0,
FEATURES             Location/Qualifiers
     source          1..898
                     /label=source [5']
                     /SerialCloner_Color=&h2F4F4F
                     /SerialCloner_Show=False
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=organism="synthetic DNA construct" - mol_type="other DNA" -  \ 
                     /SerialCloner_Desc=1..6906 \ 
     source          902..6909
                     /label=source [3']
                     /SerialCloner_Color=&h2F4F4F
                     /SerialCloner_Show=False
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=organism="synthetic DNA construct" - mol_type="other DNA" - 1..6906 \ 
     terminator      26..73
                     /label=transcription terminator for bacteriophage T7 RNA _T7 terminator
                     /SerialCloner_Color=&hF08080
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=label=T7 terminator - note="transcription terminator for bacteriophage T7 RNA  - polymerase" -  \ 
                     /SerialCloner_Desc=26..73 \ 
     CDS             complement(140..157)
                     /label=6xHis
                     /SerialCloner_Color=&hDC006B
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=codon_start=1 - product="6xHis affinity tag" - label=6xHis - translation="HHHHHH" -  \ 
                     /SerialCloner_Desc=complement(140..157) \ 
     CDS             complement(176..892)
                     /label=mammalian codon-optimized_EGFP
                     /SerialCloner_Color=&h95AE2C
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=codon_start=1 - product="enhanced GFP" - label=EGFP - note="mammalian codon-optimized" - translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL - KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD - GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYN \ 
     gene            complement(902..1705)
                     /label=mmEB1
                     /SerialCloner_Color=&hFF4500
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=product="mouse Mapre1/EB1" - label=mmEB1 -  \ 
                     /SerialCloner_Desc=complement(899..1702) \ 
     RBS             1713..1735
                     /label=efficient ribosome binding site from bacteriophage_RBS
                     /SerialCloner_Color=&hF08080
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=note="efficient ribosome binding site from bacteriophage - T7 gene 10 (Olins and Rangwala, 1989)" -  \ 
                     /SerialCloner_Desc=1710..1732 \ 
     protein_bind    1750..1774
                     /label=The lac repressor binds to the lac operator to _lac operator
                     /SerialCloner_Color=&h808000
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=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 \ 
     promoter        complement(1775..1793)
                     /label=promoter for bacteriophage T7 RNA polymerase_T7 promoter
                     /SerialCloner_Color=&hF08080
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=label=T7 promoter - note="promoter for bacteriophage T7 RNA polymerase" -  \ 
                     /SerialCloner_Desc=complement(1772..1790) \ 
     promoter        2102..2179
                     /label=lacI promoter
                     /SerialCloner_Color=&hF08080
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=gene="lacI" - label=lacI promoter -  \ 
                     /SerialCloner_Desc=2099..2176 \ 
     CDS             2180..3262
                     /label=The lac repressor binds to the lac operator to _lacI
                     /SerialCloner_Color=&h84A4C0
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=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-thiogalactopyranosid \ 
     CDS             4071..4262
                     /label=rop
                     /SerialCloner_Color=&h84A4C0
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=codon_start=1 - gene="rop" - product="Rop protein, which maintains plasmids at low copy - number" - label=rop - translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA - DELYRSCLARFGDDGENL" -  \ 
                     /SerialCloner_Desc=4068..4259 \ 
     misc_feature    4364..4506
                     /label=basis of mobility region from pBR322_bom
                     /SerialCloner_Color=&hDC143C
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=label=bom - note="basis of mobility region from pBR322" -  \ 
                     /SerialCloner_Desc=4361..4503 \ 
     rep_origin      complement(4692..5280)
                     /label=high-copy-number ColE1/pMB1/pBR322/pUC origin of _ori
                     /SerialCloner_Color=&hF08080
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=direction=LEFT - label=ori - note="high-copy-number ColE1/pMB1/pBR322/pUC origin of  - replication" -  \ 
                     /SerialCloner_Desc=complement(4689..5277) \ 
     CDS             complement(5451..6311)
                     /label=confers resistance to ampicillin, carbenicillin, and_AmpR
                     /SerialCloner_Color=&h84A4C0
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=codon_start=1 - gene="bla" - product="beta-lactamase" - label=AmpR - note="confers resistance to ampicillin, carbenicillin, and - related antibiotics" - translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI - ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQ \ 
     promoter        complement(6312..6416)
                     /label=AmpR promoter
                     /SerialCloner_Color=&hF08080
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=gene="bla" - label=AmpR promoter -  \ 
                     /SerialCloner_Desc=complement(6309..6413) \ 
     rep_origin      complement(6443..6898)
                     /label=f1 bacteriophage origin of replication; arrow _f1 ori
                     /SerialCloner_Color=&hF08080
                     /SerialCloner_Show=True
                     /SerialCloner_Protect=True
                     /SerialCloner_Arrow=True
                     /SerialCloner_Desc=direction=LEFT - label=f1 ori - note="f1 bacteriophage origin of replication; arrow  - indicates direction of (+) strand synthesis" -  \ 
                     /SerialCloner_Desc=complement(6440..6895) \ 
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa 
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt 
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcggacttgt 
      181 acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt 
      241 gatcgcgctt ctcgttgggg tctttgctca gggcggactg ggtgctcagg tagtggttgt 
      301 cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct 
      361 gcacgctgcc gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg ccgttcttct 
      421 gcttgtcggc catgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca 
      481 ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt 
      541 cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg 
      601 tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt 
      661 ggtcggggta gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc 
      721 agggcacggg cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg 
      781 catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct 
      841 cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atggtggatt 
      901 catactcttc ttgttcctcc tgtgggcccc cttcatcagg tatcacaaag ccttcatctg 
      961 tggcataaag aatatctaca atcctctgca gtacagggtc gttttccccc tcgttctcct 
     1021 ggcaaatcag ttcaatgttc cttagctttc cgaagtagaa gtctctctcc ttctccaagt 
     1081 cttcaacagt aagcttcagt actttgacct gctgcatcaa ttcagctgct tcatcatctc 
     1141 cattgcccac accaggattc tttcgcacca ttccggggcc agccttagga gctgcagtag 
     1201 tcctctgtgt tgcaatgggt ctctgtgggg ctgcagtact ggagccgaga ggtttcttcg 
     1261 gtttactcaa agctggggcg acaagagaag gagccactgc agtttcttga ccttgtctgg 
     1321 cagctacagg atcatactct tttccatcat aatttgcatc aaaaaacttc ttgaaccatt 
     1381 gaacaaattc aaaattgtcc tgaaattttc cttttactaa tttatccaca ggaattattt 
     1441 tgtcaacgcc catcctcttg aagcctgctt gtagtatttt gaagttctgg atatattcat 
     1501 gttctagctt agcttggaat ttcactttct tcaaggcaat ggatccaggg aagagcatgt 
     1561 ccataaactg acaatatgca gcccctgaac acaactgttc tatctttgtc agattcaact 
     1621 gcagagattc attgatccaa gccagcatgt catgtcgact taggttatca ctggtgactg 
     1681 acgtagagta cacattcact gccatatgta tatctccttc ttaaagttaa acaaaattat 
     1741 ttctagaggg gaattgttat ccgctcacaa ttcccctata gtgagtcgta ttaatttcgc 
     1801 gggatcgaga tctcgatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca 
     1861 ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac 
     1921 ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga 
     1981 ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg cggcggtgct caacggcctc 
     2041 aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg tcgagatccc 
     2101 ggacaccatc gaatggcgca aaacctttcg cggtatggca tgatagcgcc cggaagagag 
     2161 tcaattcagg gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg 
     2221 tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac 
     2281 gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca 
     2341 acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca 
     2401 cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt 
     2461 ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct 
     2521 tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat 
     2581 tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac 
     2641 acccatcaac agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct 
     2701 ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc 
     2761 gcgtctgcgt ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc 
     2821 ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa 
     2881 tgagggcatc gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat 
     2941 gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat atctcggtag tgggatacga 
     3001 cgataccgaa gacagctcat gttatatccc gccgttaacc accatcaaac aggattttcg 
     3061 cctgctgggg caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa 
     3121 gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa accaccctgg cgcccaatac 
     3181 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc 
     3241 ccgactggaa agcgggcagt gagcgcaacg caattaatgt aagttagctc actcattagg 
     3301 caccgggatc tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg 
     3361 cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag 
     3421 gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc tggagcgcga 
     3481 cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca cgccctcgct caagccttcg 
     3541 tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg 
     3601 ccccacgggt gcgcatgatc gtgctcctgt cgttgaggac ccggctaggc tggcggggtt 
     3661 gccttactgg ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc gactgctgct 
     3721 gcaaaacgtc tgcgacctga gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa 
     3781 gtctggaaac gcggaagtca gcgccctgca ccattatgtt ccggatctgc atcgcaggat 
     3841 gctgctggct accctgtgga acacctacat ctgtattaac gaagcgctgg cattgaccct 
     3901 gagtgatttt tctctggtcc cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt 
     3961 ccagtaaccg ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca 
     4021 tcggtatcat tacccccatg aacagaaatc ccccttacac ggaggcatca gtgaccaaac 
     4081 aggaaaaaac cgcccttaac atggcccgct ttatcagaag ccagacatta acgcttctgg 
     4141 agaaactcaa cgagctggac gcggatgaac aggcagacat ctgtgaatcg cttcacgacc 
     4201 acgctgatga gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct 
     4261 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac 
     4321 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt 
     4381 cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact 
     4441 gagagtgcac catatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc 
     4501 atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 
     4561 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 
     4621 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 
     4681 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 
     4741 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 
     4801 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 
     4861 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag 
     4921 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 
     4981 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 
     5041 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 
     5101 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg 
     5161 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 
     5221 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 
     5281 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 
     5341 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa 
     5401 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc 
     5461 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga 
     5521 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca 
     5581 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc 
     5641 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat 
     5701 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc 
     5761 attgctgcag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt 
     5821 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc 
     5881 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg 
     5941 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt 
     6001 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg 
     6061 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga 
     6121 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg 
     6181 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg 
     6241 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt 
     6301 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc 
     6361 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca 
     6421 tttccccgaa aagtgccacc tgaaattgta aacgttaata ttttgttaaa attcgcgtta 
     6481 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat 
     6541 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca 
     6601 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc 
     6661 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta 
     6721 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg 
     6781 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg 
     6841 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc 
     6901 cattcgcca
//