LOCUS       62056_18355        6754 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 6754)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6754
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             481..546
                     /codon_start=1
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags, followed by an 
                     enterokinase cleavage site"
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    763..987
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    1064..1097
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(1354..1372)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1393..1409)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1417..1433)
                     /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(1441..1471)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1486..1507)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1795..2383)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2557..3414)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3415..3519)
                     /label=AmpR promoter
     rep_origin      complement(3545..4000)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     4142..4158
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4165..4183
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(4721..5743)
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(5762..6261)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     enhancer        6333..6712
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggggggggct sgggggaggg gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg
       61 gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg
      121 tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc
      181 ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg
      241 ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg ggacggctgc
      301 cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga
      361 gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg
      421 gttattgtgc tgtctcatca ttttggcaaa gaattcctcg acggtatcga taagcttatg
      481 gactacaaag accatgacgg tgattataaa gatcatgaca tcgattacaa ggatgacgat
      541 gacaagggat cccccggggt cgacaagctt ggcgtaatca tggtcatagc tgtttcctgt
      601 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
      661 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
      721 tttccagtcg ggaaacctgt cgtgccagcg gatcagcctc gactgtgcct tctagttgcc
      781 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
      841 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
      901 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
      961 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcga
     1021 gatccactag ttctagcctc gaggctagag cggccgcgtc gagataactt cgtataatgt
     1081 atgctatacg aagttatgtt ttgttacttt atagaagaaa ttttgagttt ttgttttttt
     1141 ttaataaata aataaacata aataaattgt ttgttgaatt tattattagt atgtaagtgt
     1201 aaatataata aaacttaata tctattcaaa ttaataaata aacctcgata tacagaccga
     1261 taaaacacat gcgtcaattt tacgcatgat tatctttaac gtacgtcaca atatgattat
     1321 ctttctaggg ttaaagagct ccagcttttg ttccctttag tgagggttaa tttcgagctt
     1381 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     1441 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     1501 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     1561 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     1621 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     1681 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     1741 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1801 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1861 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1921 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1981 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2041 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2101 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2161 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2221 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2281 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2341 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2401 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2461 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2521 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2581 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     2641 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     2701 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     2761 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     2821 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     2881 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     2941 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3001 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3061 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     3121 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     3181 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     3241 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     3301 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     3361 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     3421 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     3481 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     3541 acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     3601 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     3661 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     3721 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     3781 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     3841 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     3901 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     3961 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg
     4021 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     4081 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     4141 tgtaaaacga cggccagtga attgtaatac gactcactat agggcgaatt gggtacctta
     4201 accctagaaa gatagtctgc gtaaaattga cgcatgcatt cttgaaatat tgctctctct
     4261 ttctaaatag cgcgaatccg tcgctgtgca tttaggacat ctcagtcgcc gcttggagct
     4321 cccgtgaggc gtgcttgtca atgcggtaag tgtcactgat tttgaactat aacgaccgcg
     4381 tgagtcaaaa tgacgcatga ttatctttta cgtgactttt aagatttaac tcatacgata
     4441 attatattgt tatttcatgt tctacttacg tgataactta ttatatatat attttcttgt
     4501 tatagatatc caactatgaa acattatcat aaagcaatag taaagagcct tcagtaaaga
     4561 gcaggcattt atctaatccc accccacccc cacccccgta gctccaatcc ttccattcaa
     4621 aatgtaggta ctctgttctc acccttctta acaaagtatg acaggaaaaa cttccatgtg
     4681 gacatcttta ttgtttaata gatcatcaat ttctcgacta ttcctttgcc ctcggacgag
     4741 tgctggggcg tcggtttcca ctatcggcga gtacttctac acagccatcg gtccagacgg
     4801 ccgcgcttct gcgggcgatt tgtgtacgcc cgacagtccc ggctccggat cggacgattg
     4861 cgtcgcatcg accctgcgcc caagctgcat catcgaaatt gccgtcaacc aagctctgat
     4921 agagttggtc aagaccaatg cggagcatat acgcccggag ccgcggcgat cctgcaagct
     4981 ccggatgcct ccgctcgaag tagcgcgtct gctgctccat acaagccaac cacggcctcc
     5041 agaagaagat gttggcgacc tcgtattggg aatccccgaa catcgcctcg ctccagtcaa
     5101 tgaccgctgt tatgcggcca ttgtccgtca ggacattgtt ggagccgaaa tccgcgtgca
     5161 cgaggtgccg gacttcgggg cagtcctcgg cccaaagcat cagctcatcg agagcctgcg
     5221 cgacggacgc actgacggtg tcgtccatca cagtttgcca gtgatacaca tggggatcag
     5281 caatcgcgca tatgaaatca cgccatgtag tgtattgacc gattccttgc ggtccgaatg
     5341 ggccgaaccc gctcgtctgg ctaagatcgg ccgcagcgat cgcatccatg gcctccgcga
     5401 ccggctgcag aacagcgggc agttcggttt caggcaggtc ttgcaacgtg acaccctgtg
     5461 cacggcggga gatgcaatag gtcaggctct cgctgaattc cccaatgtca agcacttccg
     5521 gaatcgggag cgcggccgat gcaaagtgcc gataaacata acgatctttg tagaaaccat
     5581 cggcgcagct atttacccgc aggacatatc cacgccctcc tacatcgaag ctgaaagcac
     5641 gagattcttc gccctccgag agctgcatca ggtcggagac gctgtcgaac ttttcgatca
     5701 gaaacttctc gacagacgtc gcggtgagtt caggcttttt catggtggcg gctggatcgg
     5761 tcgaaaggcc cggagatgag gaagaggaga acagcgcggc agacgtgcgc ttttgaagcg
     5821 tgcagaatgc cgggcctccg gaggaccttc gggcgcccgc cccgcccctg agcccgcccc
     5881 tgagcccgcc cccggaccca ccccttccca gcctctgagc ccagaaagcg aaggagcaaa
     5941 gctgctattg gccgctgccc caaaggccta cccgcttcca ttgctcagcg gtgctgtcca
     6001 tctgcacgag actagtgaga cgtgctactt ccatttgtca cgtcctgcac gacgcgagct
     6061 gcggggcggg ggggaacttc ctgactaggg gaggagtaga aggtggcgcg aaggggccac
     6121 caaagaacgg agccggttgg cgcctaccgg tggatgtgga atgtgtgcga ggccagaggc
     6181 cacttgtgta gcgccaagtg cccagcgggg ctgctaaagc gcatgctcca gactgccttg
     6241 ggaaaagcgc ctcccctacc cggtagaatt catcgatgat atcataactt cgtataatgt
     6301 atgctatacg aagttatgcg gccctcgagg tcgacattga ttattgacta gttattaata
     6361 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     6421 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     6481 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     6541 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     6601 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     6661 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt
     6721 gagccccacg ttctgcttca ctctccccat ctcc
//
