LOCUS       Exported                6736 bp ds-DNA     circular SYN 29-NOV-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6736)
  AUTHORS   MPIB
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Dec 11, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6736
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    40..64
                     /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             79..101
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             120..137
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             147..164
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             168..389
                     /codon_start=1
                     /product="human ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLR"
     CDS             396..500
                     /label=Clu(228-262)
     CDS             396..434
                     /label=Clu (alpha-Tail)
     misc_feature    435..500
                     /label=Clu (Helix7)
     CDS             507..1223
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     terminator      1295..1342
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1631..1735
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1736..2596
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2767..3355
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3541..3683
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3785..3976)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5288..6370)
                     /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(6371..6448)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
       61 ttcccctcta gaaataattt tgtttaactt taagaaggag atataccatg ggcagcagcc
      121 atcatcatca tcatcacagc agcggcctgg tgccgcgcgg cagccatatg cagatctttg
      181 tgaagaccct cactggcaaa accatcaccc ttgaggtcga gcccagtgac accattgaga
      241 atgtcaaagc caaaattcaa gacaaggagg gtatcccacc tgaccagcag cgtctgatat
      301 ttgccggcaa acagctggag gatggccgca ctctctcaga ctacaacatc cagaaagagt
      361 ccaccctgca cctggtgttg cgcctccgcg gtggtagctt gatgcccttc tctccgtacg
      421 agcccctgaa cttccacgcc atgttccagc ccttccttga gatgatacac gaggctcagc
      481 aggccatgga catccacttc ggcggcatgg tgagcaaggg cgaggagctg ttcaccgggg
      541 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
      601 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
      661 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
      721 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
      781 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
      841 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
      901 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
      961 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     1021 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     1081 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     1141 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     1201 tcggcatgga cgagctgtac aagtaagctt agatccggct gctaacaaag cccgaaagga
     1261 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     1321 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatatc ccgcaagagg
     1381 cccggcagta ccggcataac caagcctatg cctacagcat ccagggtgac ggtgccgagg
     1441 atgacgatga gcgcattgtt agatttcata cacggtgcct gactgcgtta gcaatttaac
     1501 tgtgataaac taccgcatta aagctaattc ttgaagacga aagggcctcg tgatacgcct
     1561 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     1621 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     1681 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     1741 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     1801 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     1861 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     1921 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
     1981 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     2041 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     2101 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     2161 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     2221 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
     2281 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     2341 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     2401 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     2461 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     2521 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     2581 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     2641 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     2701 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2761 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2821 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2881 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     2941 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3001 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3061 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3121 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3181 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3241 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3301 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3361 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     3421 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     3481 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     3541 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc
     3601 actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc
     3661 tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac
     3721 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     3781 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat
     3841 cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga
     3901 gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt
     3961 tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga
     4021 taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt
     4081 tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa
     4141 tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc
     4201 agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt
     4261 ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg
     4321 ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag
     4381 taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc
     4441 gtggccagga cccaacgctg cccgagatgc gccgcgtgcg gctgctggag atggcggacg
     4501 cgatggatat gttctgccaa gggttggttt gcgcattcac agttctccgc aagaattgat
     4561 tggctccaat tcttggagtg gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt
     4621 cgaggtggcc cggctccatg caccgcgacg caacgcgggg aggcagacaa ggtatagggc
     4681 ggcgcctaca atccatgcca acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt
     4741 gacgatcagc ggtccagtga tcgaagttag gctggtaaga gccgcgagcg atccttgaag
     4801 ctgtccctga tggtcgtcat ctacctgcct ggacagcatg gcctgcaacg cgggcatccc
     4861 gatgccgccg gaagcgagaa gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa
     4921 cgccagcaag acgtagccca gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc
     4981 gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc
     5041 gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa
     5101 aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat
     5161 aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     5221 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
     5281 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5341 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca
     5401 ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca
     5461 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg
     5521 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa
     5581 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
     5641 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
     5701 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
     5761 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
     5821 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt
     5881 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg
     5941 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa
     6001 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga
     6061 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt
     6121 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa
     6181 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct
     6241 ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca
     6301 ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta
     6361 ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc
     6421 gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc
     6481 tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat
     6541 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
     6601 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     6661 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     6721 ccggcgtaga ggatcg
//
