LOCUS       62056_3340        7559 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 7559)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7559
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..657
                     /label=chicken beta-actin promoter
     intron          658..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and
                     rabbit beta-globin"
     polyA_signal    1819..2043
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    complement(2128..2245)
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     protein_bind    2291..2324
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(2581..2599)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2620..2636)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2644..2660)
                     /label=lac operator
                     /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)."
     promoter        complement(2668..2698)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2713..2734)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3022..3610)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3784..4641)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4642..4746)
                     /label=AmpR promoter
     rep_origin      complement(4772..5227)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     5369..5385
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5392..5410
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(5948..6970)
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(6989..7488)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     promoter        7557..7559
                     /label=CAG
                     /note="CMV early enhancer fused to modified chicken 
                     beta-actin promoter"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc
      541 ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct
      601 tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga
      661 gtcgctgcgt tgccttcgcc ccgtgccccg ctccgcgccg cctcgcgccg cccgccccgg
      721 ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc
      781 tgtaattagc gcttggttta atgacggctc gtttcttttc tgtggctgcg tgaaagcctt
      841 aaagggctcc gggagggccc tttgtgcggg ggggagcggc tcggggggtg cgtgcgtgtg
      901 tgtgtgcgtg gggagcgccg cgtgcggccc gcgctgcccg gcggctgtga gcgctgcggg
      961 cgcggcgcgg ggctttgtgc gctccgcgtg tgcgcgaggg gagcgcggcc gggggcggtg
     1021 ccccgcggtg cgggggggct gcgaggggaa caaaggctgc gtgcggggtg tgtgcgtggg
     1081 ggggtgagca gggggtgtgg gcgcggcggt cgggctgtaa cccccccctg cacccccctc
     1141 cccgagttgc tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg
     1201 ggctcgccgt gccgggcggg gggtggcggc aggtgggggt gccgggcggg gcggggccgc
     1261 ctcgggccgg ggagggctcg ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc
     1321 gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac
     1381 ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc accccctcta
     1441 gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg gagggccttc
     1501 gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt ccgcgggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcctcgac ggtatcgata
     1741 agctatcaca agtttgtaca ttcgaaggat ccacgcgtct cgagttaacg gtacctctag
     1801 agaattcgct agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     1861 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     1921 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     1981 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     2041 tggcttctga ggcggttaat taaaataaaa ctagagcggg cccggtacta accaaactgg
     2101 atctctgctg tccctgtaat aaacccgaaa attttgaatt tttgtaattt gtttttgtaa
     2161 ttctttagtt tgtatgtctg ttgctattat gtctactatt ctttcccctg cactgtaccc
     2221 cccaatcccc ccttttcttt taaaattgtg gatgaatact gccatttgtc tgcagagcgg
     2281 ccgcgtcgag ataacttcgt ataatgtatg ctatacgaag ttatgttttg ttactttata
     2341 gaagaaattt tgagtttttg ttttttttta ataaataaat aaacataaat aaattgtttg
     2401 ttgaatttat tattagtatg taagtgtaaa tataataaaa cttaatatct attcaaatta
     2461 ataaataaac ctcgatatac agaccgataa aacacatgcg tcaattttac gcatgattat
     2521 ctttaacgta cgtcacaata tgattatctt tctagggtta aagagctcca gcttttgttc
     2581 cctttagtga gggttaattt cgagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     2641 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     2701 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     2761 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     2821 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2881 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2941 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3001 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3061 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3121 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3181 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3241 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3301 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3361 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3421 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3481 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3541 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3601 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3661 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3721 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3781 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3841 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3901 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3961 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4021 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4081 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4141 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4201 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4261 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4321 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4381 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4441 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4501 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4561 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4621 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4681 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4741 tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa
     4801 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     4861 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     4921 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     4981 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     5041 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     5101 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     5161 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     5221 ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     5281 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     5341 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac
     5401 tcactatagg gcgaattggg taccttaacc ctagaaagat agtctgcgta aaattgacgc
     5461 atgcattctt gaaatattgc tctctctttc taaatagcgc gaatccgtcg ctgtgcattt
     5521 aggacatctc agtcgccgct tggagctccc gtgaggcgtg cttgtcaatg cggtaagtgt
     5581 cactgatttt gaactataac gaccgcgtga gtcaaaatga cgcatgatta tcttttacgt
     5641 gacttttaag atttaactca tacgataatt atattgttat ttcatgttct acttacgtga
     5701 taacttatta tatatatatt ttcttgttat agatatccaa ctatgaaaca ttatcataaa
     5761 gcaatagtaa agagccttca gtaaagagca ggcatttatc taatcccacc ccacccccac
     5821 ccccgtagct ccaatccttc cattcaaaat gtaggtactc tgttctcacc cttcttaaca
     5881 aagtatgaca ggaaaaactt ccatgtggac atctttattg tttaatagat catcaatttc
     5941 tcgactattc ctttgccctc ggacgagtgc tggggcgtcg gtttccacta tcggcgagta
     6001 cttctacaca gccatcggtc cagacggccg cgcttctgcg ggcgatttgt gtacgcccga
     6061 cagtcccggc tccggatcgg acgattgcgt cgcatcgacc ctgcgcccaa gctgcatcat
     6121 cgaaattgcc gtcaaccaag ctctgataga gttggtcaag accaatgcgg agcatatacg
     6181 cccggagccg cggcgatcct gcaagctccg gatgcctccg ctcgaagtag cgcgtctgct
     6241 gctccataca agccaaccac ggcctccaga agaagatgtt ggcgacctcg tattgggaat
     6301 ccccgaacat cgcctcgctc cagtcaatga ccgctgttat gcggccattg tccgtcagga
     6361 cattgttgga gccgaaatcc gcgtgcacga ggtgccggac ttcggggcag tcctcggccc
     6421 aaagcatcag ctcatcgaga gcctgcgcga cggacgcact gacggtgtcg tccatcacag
     6481 tttgccagtg atacacatgg ggatcagcaa tcgcgcatat gaaatcacgc catgtagtgt
     6541 attgaccgat tccttgcggt ccgaatgggc cgaacccgct cgtctggcta agatcggccg
     6601 cagcgatcgc atccatggcc tccgcgaccg gctgcagaac agcgggcagt tcggtttcag
     6661 gcaggtcttg caacgtgaca ccctgtgcac ggcgggagat gcaataggtc aggctctcgc
     6721 tgaattcccc aatgtcaagc acttccggaa tcgggagcgc ggccgatgca aagtgccgat
     6781 aaacataacg atctttgtag aaaccatcgg cgcagctatt tacccgcagg acatatccac
     6841 gccctcctac atcgaagctg aaagcacgag attcttcgcc ctccgagagc tgcatcaggt
     6901 cggagacgct gtcgaacttt tcgatcagaa acttctcgac agacgtcgcg gtgagttcag
     6961 gctttttcat ggtggcggct ggatcggtcg aaaggcccgg agatgaggaa gaggagaaca
     7021 gcgcggcaga cgtgcgcttt tgaagcgtgc agaatgccgg gcctccggag gaccttcggg
     7081 cgcccgcccc gcccctgagc ccgcccctga gcccgccccc ggacccaccc cttcccagcc
     7141 tctgagccca gaaagcgaag gagcaaagct gctattggcc gctgccccaa aggcctaccc
     7201 gcttccattg ctcagcggtg ctgtccatct gcacgagact agtgagacgt gctacttcca
     7261 tttgtcacgt cctgcacgac gcgagctgcg gggcgggggg gaacttcctg actaggggag
     7321 gagtagaagg tggcgcgaag gggccaccaa agaacggagc cggttggcgc ctaccggtgg
     7381 atgtggaatg tgtgcgaggc cagaggccac ttgtgtagcg ccaagtgccc agcggggctg
     7441 ctaaagcgca tgctccagac tgccttggga aaagcgcctc ccctacccgg tagaattcat
     7501 cgatgatatc ataacttcgt ataatgtatg ctatacgaag ttatgcggcc ctcgaggtc
//
