LOCUS       62056_21985        4828 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 4828)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        142..172
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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)."
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             298..1014
                     /codon_start=1
                     /label=EBFP2
                     /note="enhanced blue variant of GFP (Ai et al., 2007)"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     protein_bind    1018..1051
                     /label=loxP511
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are 
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     polyA_signal    1052..1276
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        1277..1776
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             1786..2157
                     /codon_start=1
                     /label=BleoR
                     /note="antibiotic-binding protein"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     primer_bind     complement(2432..2448)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2922..3026
                     /label=AmpR promoter
     CDS             3027..3884
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4058..4646
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttca
      241 tatgagggca gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc cggccctatg
      301 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
      361 gacgtaaacg gccacaagtt cagcgtgagg ggcgagggcg agggcgatgc caccaacggc
      421 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
      481 gtgaccaccc tgagccacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag
      541 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
      601 aaggacgacg gcacctacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
      661 aaccgcatcg agctgaaggg cgtcgacttc aaggaggacg gcaacatcct ggggcacaag
      721 ctggagtaca acttcaacag ccacaacatc tatatcatgg ccgtcaagca gaagaacggc
      781 atcaaggtga acttcaagat ccgccacaac gtggaggacg gcagcgtgca gctcgccgac
      841 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga cagccactac
      901 ctgagcaccc agtccgtgct gagcaaagac cccaacgaga agcgcgatca catggtcctg
      961 ctggagttcc gcaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaata
     1021 acttcgtata atgtatacta tacgaagtta tctgtgcctt ctagttgcca gccatctgtt
     1081 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     1141 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     1201 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     1261 gcggtgggct ctatgggggt aggggaggcg cttttcccaa ggcagtctgg agcatgcgct
     1321 ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc
     1381 acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc gccaccttct
     1441 actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg caggacgtga
     1501 caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc tgagcaatgg
     1561 aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct
     1621 cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg
     1681 ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc
     1741 gcgctgttct cctcttcctc atctccgggc ctttcgaggc tcgagatggc caagttgacc
     1801 agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac
     1861 cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac
     1921 gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg
     1981 gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac
     2041 ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag
     2101 ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga
     2161 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     2221 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     2281 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     2341 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggataac ttcgtataat
     2401 gtatactata cgaagttatg agctcgaatt cactggccgt cgttttacaa cgtcgtgact
     2461 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     2521 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     2581 gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     2641 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     2701 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     2761 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     2821 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     2881 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     2941 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3001 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     3061 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3121 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     3181 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     3241 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     3301 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     3361 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3421 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     3481 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     3541 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     3601 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     3661 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3721 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     3781 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     3841 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     3901 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     3961 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4021 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4081 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4141 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4201 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4261 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4321 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4381 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4441 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4501 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4561 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4621 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4681 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     4741 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     4801 gcgagtcagt gagcgaggaa gcggaaga
//
