LOCUS       40924_7221        6919 bp DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression of Cre recombinase driven by the human 
            cytomegalovirus IE promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6919)
  AUTHORS   Sauer B, Henderson N
  TITLE     Targeted insertion of exogenous DNA into the eukaryotic genome by 
            the Cre recombinase.
  JOURNAL   New Biol. 1990 May . 2(5):441-9.
  PUBMED    2288914
REFERENCE   2  (bases 1 to 6919)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6919)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "New Biol. 
            1990 May . 2(5):441-9."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6919
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(565..581)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(589..605)
                     /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(613..643)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(658..679)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     primer_bind     complement(796..813)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(967..1555)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1729..2586)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2587..2691)
                     /label=AmpR promoter
     primer_bind     2759..2777
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     primer_bind     complement(2815..2837)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2937..2956
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     primer_bind     3165..3181
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        3267..3646
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3647..3850
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     primer_bind     3847..3871
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3991..5019
                     /codon_start=1
                     /label=Cre
                     /note="site-specific recombinase"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     RBS             6674..6682
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
ORIGIN
        1 cactattgca tatgccatca agattttgct gaaaggaccc tgataaagct gtctcctgtg
       61 aggctttgcc agtgcccagc aaacacagaa gtggatgctc acagtcagct attggatgga
      121 acacagggcc cccaatggag gagctagaga aagtacccaa ggagctaaag ggatctgcaa
      181 ccctataggt ggaacagcaa tctgaactaa ccagtacccc ctgaactcgt gtatctagct
      241 tcgtatgtag cagaagatgg cctagttggc catcactggg aagagaggac ccttggtcta
      301 gcaaactata tgccccagtt caggggaaag ccagggccaa gaagtgggag tgagtgggta
      361 ggggagcggg gtgggggtag gggtggggta taggggactt ttgggatagc atttgaaata
      421 taaacgaaga aaatatccaa aaaaaaaaag atggaaaaaa aagtgataaa gagcgtgggt
      481 cctgcctgta atcaagatat ttgagatgta gcagcaggaa ggccaggaat tggtcgaggg
      541 taccgagctc gaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct
      601 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
      661 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
      721 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
      781 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
      841 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
      901 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
      961 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1021 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1081 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1141 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1201 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1261 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1321 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     1381 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     1441 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     1501 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1561 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     1621 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     1681 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     1741 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     1801 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     1861 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     1921 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     1981 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     2041 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     2101 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     2161 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     2221 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     2281 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     2341 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     2401 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     2461 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     2521 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     2581 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     2641 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     2701 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     2761 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     2821 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     2881 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     2941 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     3001 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     3061 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     3121 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     3181 tgccaagctt ggcccattgc atacgttgta tccatatcat aatatgtaca tttatattgg
     3241 ctcatgtcca acattaccgc catgttgaca ttgattattg actagttatt aatagtaatc
     3301 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     3361 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     3421 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     3481 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     3541 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     3601 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     3661 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     3721 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     3781 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     3841 aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga
     3901 cctccataga agacaccgac tctagaggat cccctcgagg ggcagagccg atcctgtaca
     3961 ctttacttaa aaccattatc tgagtgtgaa atgtccaatt tactgaccgt acaccaaaat
     4021 ttgcctgcat taccggtcga tgcaacgagt gatgaggttc gcaagaacct gatggacatg
     4081 ttcagggatc gccaggcgtt ttctgagcat acctggaaaa tgcttctgtc cgtttgccgg
     4141 tcgtgggcgg catggtgcaa gttgaataac cggaaatggt ttcccgcaga acctgaagat
     4201 gttcgcgatt atcttctata tcttcaggcg cgcggtctgg cagtaaaaac tatccagcaa
     4261 catttgggcc agctaaacat gcttcatcgt cggtccgggc tgccacgacc aagtgacagc
     4321 aatgctgttt cactggttat gcggcggatc cgaaaagaaa acgttgatgc cggtgaacgt
     4381 gcaaaacagg ctctagcgtt cgaacgcact gatttcgacc aggttcgttc actcatggaa
     4441 aatagcgatc gctgccagga tatacgtaat ctggcatttc tggggattgc ttataacacc
     4501 ctgttacgta tagccgaaat tgccaggatc agggttaaag atatctcacg tactgacggt
     4561 gggagaatgt taatccatat tggcagaacg aaaacgctgg ttagcaccgc aggtgtagag
     4621 aaggcactta gcctgggggt aactaaactg gtcgagcgat ggatttccgt ctctggtgta
     4681 gctgatgatc cgaataacta cctgttttgc cgggtcagaa aaaatggtgt tgccgcgcca
     4741 tctgccacca gccagctatc aactcgcgcc ctggaaggga tttttgaagc aactcatcga
     4801 ttgatttacg gcgctaagga tgactctggt cagagatacc tggcctggtc tggacacagt
     4861 gcccgtgtcg gagccgcgcg agatatggcc cgcgctggag tttcaatacc ggagatcatg
     4921 caagctggtg gctggaccaa tgtaaatatt gtcatgaact atatccgtaa cctggatagt
     4981 gaaacagggg caatggtgcg cctgctggaa gatggcgatt agccattaac gcgtaaatga
     5041 ttgctataat tatttgatat ttatggtgac atatgagaaa ggatttcaac atcgacggaa
     5101 aatatgtagt gctgtctgta agcactaata ttcagtcgcc agccgtcatt gtcactgtaa
     5161 agctgagcga tagaatgcct gatattgact caatatccgt tgcgtttcct gtcaaaagta
     5221 tgcgtagtgc tgaacatttc gtgatgaatg ccaccgagga agaagcacgg cgcggttttg
     5281 ctaaagtgat gtctgagttt ggcgaactct tgggtaaggt tggaattgtc gagggatctc
     5341 ggaatggacc ccaactgctc ctgctccacc ggtaagactc ccgatccttg gtctttagaa
     5401 taccaagttg ggaccgcaga gcggaatccc cgagttgtag aggcttggcg ggaataggca
     5461 cctttagttg gcgattcatt ccggttcttt ctagaatccg ctcttgcaaa agccttcatt
     5521 agttacgagt attgtcgaac gggtcctttg gcggggttgg ggctaggatt tagacgcgca
     5581 aatgtccggt tcctgatcac ccagttagtg gggacatctg ggttgagtcc caggcattac
     5641 taaacttact gtgaattgct tgaattaaga aagaggtgaa ggacctttat gtcttgggac
     5701 tcaaagacat aatccctgac ttaacctgtg aggagaaaag tggggctagg ctccctgcag
     5761 ctccgaggag gacttagtga actgagccgg gactcgtggt gttggccact gctgtaatgc
     5821 tgcctccctc atgctgtctt ctttctcctc cccaggcggc tcctgcactt gcaccagctc
     5881 ctgcgcctgc aagaactgca agtgcacctc ctgcaagaag agtgagttgg gacaccttgg
     5941 gtggcggcta aggctagggg cggggaactc ctacaaaact ggctctgaga aatgtccttt
     6001 gcttcccgga ggccattgta ttgtctcggg gacagaacta tacagagaac tatttaaaaa
     6061 aaccgaggtc ttctctgttg gggacaggaa gcagaggtct tcagccaggc tgacctcttc
     6121 ctcctccttt ctaggctgct gctcctgctg tcccgtgggc tgctccaaat gtgcccaggg
     6181 ctgtgtctgc aaaggcgccg cggacaagtg cacgtgctgt gcctgatgtg acgaacagcg
     6241 ctgccaccac gtgtaaatag tatcggacca acccagcgtc ttcctataca gttccaccct
     6301 gtttactaaa cccccgtttt ctaccgagta cgtgaataat aaaagcctgt ttgagtctaa
     6361 ctctggtttt cttggtgtgg tttggcaata agaaactggg gtgacttgat agtctgggga
     6421 tctggttttg gaccccctcg tgcctttacc tcccgcctct ggccctcaca gaggggtaat
     6481 gtctttgggt aaagccaagc tatcccataa gcttcctcat ggaaaacagc tgtgtgcctg
     6541 gtcaggtctt gtgttagggg aagggaggga gggagggtgg tcctggaaga cttgatgtcc
     6601 catcctggaa gcacagaaaa tcttatcatt tatattctct ttggattctg gtttcctctc
     6661 acattgttgg gtataaggag gtgggtatga aaattaacat agggtgtctc gtatgctagg
     6721 cagggactaa cactgaacta gattccagag gggctttctt ggctgttttg acacaagttg
     6781 cttaccttgt cctagaactc attctgcagt ccagtctgac cttgagtttt tgattctcct
     6841 gcctcagctt cccaagtacc tgggattaca ggagtgactg tgtcaccagg ccatctccta
     6901 ttcagccacc aaatgcaga
//
