LOCUS       62056_12915        4979 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4979)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4979
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(4..861)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1031..1619
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     enhancer        1794..2097
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2098..2301
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        2347..2365
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             2443..2514
                     /codon_start=1
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags"
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     misc_feature    2550..3133
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             3134..3850
                     /codon_start=1
                     /label=hrGFP
                     /note="humanized Renilla green fluorescent protein"
                     /translation="MVSKQILKNTGLQEIMSFKVNLEGVVNNHVFTMEGCGKGNILFGN
                     QLVQIRVTKGAPLPFAFDILSPAFQYGNRTFTKYPEDISDFFIQSFPAGFVYERTLRYE
                     DGGLVEIRSDINLIEEMFVYRVEYKGRNFPNDGPVMKKTITGLQPSFEVVYMNDGVLVG
                     QVILVYRLNSGKFYSCHMRTLMKSKGVVKDFPEYHFIQHRLEKTYVEDGGFVEQHETAI
                     AQLTSLGKPLGSLHEWV"
     promoter        complement(3884..3902)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    4176..4297
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4304..4759)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4786..4888
                     /label=AmpR promoter
     protein_bind    4905..4938
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
       61 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
      121 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
      181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
      241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
      301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
      361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
      421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
      481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
      541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
      601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
      661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
      721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
      781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
      841 acggaaatgt tgaatactca tactcttcct caggttactc atatatactt tagattgatt
      901 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
      961 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     1021 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     1081 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     1141 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     1201 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     1261 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     1321 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     1381 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     1441 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     1501 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     1561 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     1621 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     1681 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat
     1741 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     1801 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     1861 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     1921 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     1981 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     2041 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
     2101 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
     2161 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     2221 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
     2281 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg attacgccaa
     2341 gctcgaaatt aaccctcact aaagggaaca aaagctggag ctccaccgcg gtggcggccg
     2401 ctctagcccg ggcggatccg aattcgcatg cgtcgactcg aggactacaa ggatgacgat
     2461 gacaaggatt acaaagacga cgatgataag gactataagg atgatgacga caaataatag
     2521 caattcctcg acgactgcat agggttaccc ccctctccct cccccccccc taacgttact
     2581 ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata
     2641 ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt
     2701 cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa
     2761 gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag
     2821 cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca
     2881 cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc
     2941 aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat
     3001 tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta
     3061 aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat
     3121 aatggccaca accatggtga gcaagcagat cctgaagaac accggcctgc aggagatcat
     3181 gagcttcaag gtgaacctgg agggcgtggt gaacaaccac gtgttcacca tggagggctg
     3241 cggcaagggc aacatcctgt tcggcaacca gctggtgcag atccgcgtga ccaagggcgc
     3301 ccccctgccc ttcgccttcg acatcctgag ccccgccttc cagtacggca accgcacctt
     3361 caccaagtac cccgaggaca tcagcgactt cttcatccag agcttccccg ccggcttcgt
     3421 gtacgagcgc accctgcgct acgaggacgg cggcctggtg gagatccgca gcgacatcaa
     3481 cctgatcgag gagatgttcg tgtaccgcgt ggagtacaag ggccgcaact tccccaacga
     3541 cggccccgtg atgaagaaga ccatcaccgg cctgcagccc agcttcgagg tggtgtacat
     3601 gaacgacggc gtgctggtgg gccaggtgat cctggtgtac cgcctgaaca gcggcaagtt
     3661 ctacagctgc cacatgcgca ccctgatgaa gagcaagggc gtggtgaagg acttccccga
     3721 gtaccacttc atccagcacc gcctggagaa gacctacgtg gaggacggcg gcttcgtgga
     3781 gcagcacgag accgccatcg cccagctgac cagcctgggc aagcccctgg gcagcctgca
     3841 cgagtgggtg taatagggta ccaggtaagt gtacccaatt cgccctatag tgagtcgtat
     3901 tacaattcac tcgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggagatcc
     3961 aatttttaag tgtataatgt gttaaactac tgattctaat tgtttgtgta ttttagattc
     4021 acagtcccaa ggctcatttc aggcccctca gtcctcacag tctgttcatg atcataatca
     4081 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     4141 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     4201 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4261 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat
     4321 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     4381 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     4441 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     4501 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     4561 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     4621 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     4681 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     4741 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4801 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4861 atgcttcaat aatattgaaa aaggaagaat cctgaggccg ggccataact tcgtataatg
     4921 tatgctatac gaagttatcc atgggccccc cctcgacatg agtaaacttg gtctgacag
//
