LOCUS       Exported                8977 bp DNA     circular SYN 12-MAY-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8977)
  AUTHORS   Universitaetsklinikum Halle
  TITLE     Direct Submission
  JOURNAL   Exported Jun 3, 2022 from SnapGene Viewer 6.0.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8977
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    235..256
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        271..301
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    309..325
                     /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     333..349
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        370..388
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        416..642
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             643..823
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    870..995
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1488..1721
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    2217..2334
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2368..2774
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     CDS             2837..3865
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DG"
     misc_feature    3895..4481
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             4482..5183
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     misc_feature    5418..5821
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             5911..6144
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6216..6337
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      6377..6512
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(6533..6551)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6561..6577)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6719..7174
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        7200..7304
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7305..8165
                     /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"
     rep_origin      8336..8924
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
       61 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
      121 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
      181 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
      241 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      301 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
      361 caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag cttaatgtag
      421 tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca acatgcctta
      481 caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta cgatcgtgcc
      541 ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga attgccgcat
      601 tgcagagata ttgtatttaa gtgcctagct cgatacataa acgggtctct ctggttagac
      661 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
      721 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
      781 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg
      841 acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag
      901 cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg
      961 gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat
     1021 cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata
     1081 tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat
     1141 cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag
     1201 aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga
     1261 taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca
     1321 ccgcacagca agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg
     1381 agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc
     1441 aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc
     1501 cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta
     1561 caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt
     1621 gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga
     1681 atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct
     1741 ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg
     1801 gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca
     1861 agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa
     1921 ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg
     1981 tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt
     2041 gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc
     2101 cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag
     2161 agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg gttaactttt
     2221 aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca
     2281 acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttatcgat
     2341 cacgagacta gcctcgagct agctgcagta acgccatttt gcaaggcatg gaaaaatacc
     2401 aaaccaagaa tagagaagtt cagatcaagg gcgggtacat gaaaatagct aacgttgggc
     2461 caaacaggat atctgcggtg agcagtttcg gccccggccc ggggccaaga acagatggtc
     2521 accgcagttt cggccccggc ccgaggccaa gaacagatgg tccccagata tggcccaacc
     2581 ctcagcagtt tcttaagacc catcagatgt ttccaggctc ccccaaggac ctgaaatgac
     2641 cctgcgcctt atttgaatta accaatcagc ctgcttctcg cttctgttcg cgcgcttctg
     2701 cttcccgagc tctataaaag agctcacaac ccctcactcg gcgcgccagt cctccgacag
     2761 actgagtcgg ccggtcgaat caagcttatc gataccgtcg acggatccac cggtactagt
     2821 atactccgga gccaccatgt ccaatttact gaccgtacac caaaatttgc ctgcattacc
     2881 ggtcgatgca acgagtgatg aggttcgcaa gaacctgatg gacatgttca gggatcgcca
     2941 ggcgttttct gagcatacct ggaaaatgct tctgtccgtt tgccggtcgt gggcggcatg
     3001 gtgcaagttg aataaccgga aatggtttcc cgcagaacct gaagatgttc gcgattatct
     3061 tctatatctt caggcgcgcg gtctggcagt aaaaactatc cagcaacatt tgggccagct
     3121 aaacatgctt catcgtcggt ccgggctgcc acgaccaagt gacagcaatg ctgtttcact
     3181 ggttatgcgg cggatccgaa aagaaaacgt tgatgccggt gaacgtgcaa aacaggctct
     3241 agcgttcgaa cgcactgatt tcgaccaggt tcgttcactc atggaaaata gcgatcgctg
     3301 ccaggatata cgtaatctgg catttctggg gattgcttat aacaccctgt tacgtatagc
     3361 cgaaattgcc aggatcaggg ttaaagatat ctcacgtact gacggtggga gaatgttaat
     3421 ccatattggc agaacgaaaa cgctggttag caccgcaggt gtagagaagg cacttagcct
     3481 gggggtaact aaactggtcg agcgatggat ttccgtctct ggtgtagctg atgatccgaa
     3541 taactacctg ttttgccggg tcagaaaaaa tggtgttgcc gcgccatctg ccaccagcca
     3601 gctatcaact cgcgccctgg aagggatttt tgaagcaact catcgattga tttacggcgc
     3661 taaggatgac tctggtcaga gatacctggc ctggtctgga cacagtgccc gtgtcggagc
     3721 cgcgcgagat atggcccgcg ctggagtttc aataccggag atcatgcaag ctggtggctg
     3781 gaccaatgta aatattgtca tgaactatat ccgtaacctg gatagtgaaa caggggcaat
     3841 ggtgcgcctg ctggaagatg gctgaacgcg tatgcatgag taactgagga tccgcccctc
     3901 tccctncccc ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt
     3961 tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc
     4021 tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca
     4081 aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac
     4141 gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg
     4201 ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt
     4261 gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct
     4321 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg
     4381 ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt
     4441 ggttttcctt tgaaaaacac gatgataata tggccacaac catggtgagc aagggcgagg
     4501 aggtcatcaa agagttcatg cgcttcaagg tgcgcatgga gggctccatg aacggccacg
     4561 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc
     4621 tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc ccccagttca
     4681 tgtacggctc caaggcgtac gtgaagcacc ccgccgacat ccccgattac aagaagctgt
     4741 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggtctggtga
     4801 ccgtgaccca ggactcctcc ctgcaggacg gcacgctgat ctacaaggtg aagatgcgcg
     4861 gcaccaactt cccccccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct
     4921 ccaccgagcg cctgtacccc cgcgacggcg tgctgaaggg cgagatccac caggccctga
     4981 agctgaagga cggcggccac tacctggtgg agttcaagac catctacatg gccaagaagc
     5041 ccgtgcaact gcccggctac tactacgtgg acaccaagct ggacatcacc tcccacaacg
     5101 aggactacac catcgtggaa cagtacgagc gctccgaggg ccgccaccac ctgttcctgt
     5161 acggcatgga cgagctgtac aagtaaagcg gccgcgtcga cggatccccc gggctgcagg
     5221 aattcgagca tcttaccgcc atttattccc atatttgttc tgtttttctt gatttgggta
     5281 tacatttaaa tgttaataaa acaaaatggt ggggcaatca tttacatttt tagggatatg
     5341 taattactag ttcaggtgta ttgccacaag acaaacatgt taagaaactt tcccgttatt
     5401 tacgctctgt tcctgttaat caacctctgg attacaaaat ttgtgaaaga ttgactgata
     5461 ttcttaacta tgttgctcct tttacgctgt gtggatatgc tgctttaatg cctctgtatc
     5521 atgctattgc ttcccgtacg gctttcgttt tctcctcctt gtataaatcc tggttgctgt
     5581 ctctttatga ggagttgtgg cccgttgtcc gtcaacgtgg cgtggtgtgc tctgtgtttg
     5641 ctgacgcaac ccccactggc tggggcattg ccaccacctg tcaactcctt tctgggactt
     5701 tcgctttccc cctcccgatc gccacggcag aactcatcgc cgcctgcctt gcccgctgct
     5761 ggacaggggc taggttgctg ggcactgata attccgtggt gttgtcgggg aagctgacgt
     5821 cctttcgaat tcgatatcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     5881 ttagccactt tttaaaagaa aaggggggac tggaagggct acgtaactcc caacgaagac
     5941 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     6001 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     6061 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     6121 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
     6181 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
     6241 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6301 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
     6361 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     6421 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     6481 ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt
     6541 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     6601 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     6661 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     6721 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     6781 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     6841 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     6901 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     6961 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     7021 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     7081 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     7141 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     7201 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     7261 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     7321 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     7381 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     7441 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     7501 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     7561 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     7621 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     7681 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     7741 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     7801 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     7861 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     7921 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     7981 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     8041 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     8101 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     8161 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     8221 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     8281 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     8341 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     8401 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     8461 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     8521 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     8581 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     8641 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     8701 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     8761 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     8821 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     8881 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     8941 cctttttacg gttcctggcc ttttgctggc cttttgc
//
