LOCUS       62056_7540        7854 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 7854)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7854
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    complement(124..258)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        645..663
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(1009..4053)
                     /codon_start=1
                     /label=lacZ
                     /note="beta-galactosidase"
                     /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS
                     LNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNPP
                     FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF
                     DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR
                     FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV
                     TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP
                     LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL
                     YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA
                     NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS
                     LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP
                     WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL
                     FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ
                     PNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ
                     SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA
                     ALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP
                     ARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGLR
                     CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD
                     SWSPSVSAELQLSAGRYHYQLVWCQK"
     promoter        complement(4260..4278)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     promoter        complement(4394..4597)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     enhancer        complement(4598..4901)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    complement(4931..4947)
                     /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(4955..4986)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     rep_origin      complement(5449..6037)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    complement(6115..6148)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(6253..7110)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      complement(7398..7853)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
       61 ttacgccagc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagg
      121 atcgatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
      181 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
      241 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
      301 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat
      361 gatcatgaac agactgtgag gactgagggg cctgaaatga gccttgggac tgtgaatcta
      421 aaatacacaa acaattagaa tcactagctc ctgtgtataa tattttcata aatcatactc
      481 agtaagcaaa actctcaagc agcaagcata tgcagctagt ttaacacatt atacacttaa
      541 aaattttata tttaccttag agctttaaat ctctgtaggt agtttgtcca attatgtcac
      601 accacagaag taaggttcct tcacaaagat cccaagctag cttataatac gactcactat
      661 agggagagag ctatgacgtc gcatgcacgc gtaagcttgg gcccctcgag ggatcctcta
      721 gagcggccgc ctagagtcga ggccgagttt gtcagaaagc agaccaaaca gcggttggaa
      781 taatagcgag aacagagaaa tagcggcaaa aataataccc gtatcacttt tgctgatatg
      841 gttgatgtca tgtagccaaa tcgggaaaaa cgggaagtag gctcccatga taaaaaagta
      901 aaagaaaaag aataaaccga acatccaaaa gtttgtgttt tttaaatagt acataatgga
      961 tttccttacg cgaaatacgg gcagacatgg cctgcccggt tattattatt tttgacacca
     1021 gaccaactgg taatggtagc gaccggcgct cagctgtaat tccgccgata ctgacgggct
     1081 ccaggagtcg tcgccaccaa tccccatatg gaaaccgtcg atattcagcc atgtgccttc
     1141 ttccgcgtgc agcagatggc gatggctggt ttccatcagt tgctgttgac tgtagcggct
     1201 gatgttgaac tggaagtcgc cgcgccactg gtgtgggcca taattcaatt cgcgcgtccc
     1261 gcagcgcaga ccgttttcgc tcgggaagac gtacggggta tacatgtctg acaatggcag
     1321 atcccagcgg tcaaaacagg cggcagtaag gcggtcggga tagttttctt gcggccctaa
     1381 tccgagccag tttacccgct ctgctacctg cgccagctgg cagttcaggc caatccgcgc
     1441 cggatgcggt gtatcgctcg ccacttcaac atcaacggta atcgccattt gaccactacc
     1501 atcaatccgg taggttttcc ggctgataaa taaggttttc ccctgatgct gccacgcgtg
     1561 agcggtcgta atcagcaccg catcagcaag tgtatctgcc gtgcactgca acaacgctgc
     1621 ttcggcctgg taatggcccg ccgccttcca gcgttcgacc caggcgttag ggtcaatgcg
     1681 ggtcgcttca cttacgccaa tgtcgttatc cagcggtgca cgggtgaact gatcgcgcag
     1741 cggcgtcagc agttgttttt tatcgccaat ccacatctgt gaaagaaagc ctgactggcg
     1801 gttaaattgc caacgcttat tacccagctc gatgcaaaaa tccatttcgc tggtggtcag
     1861 atgcgggatg gcgtgggacg cggcggggag cgtcacactg aggttttccg ccagacgcca
     1921 ctgctgccag gcgctgatgt gcccggcttc tgaccatgcg gtcgcgttcg gttgcactac
     1981 gcgtactgtg agccagagtt gcccggcgct ctccggctgc ggtagttcag gcagttcaat
     2041 caactgttta ccttgtggag cgacatccag aggcacttca ccgcttgcca gcggcttacc
     2101 atccagcgcc accatccagt gcaggagctc gttatcgcta tgacggaaca ggtattcgct
     2161 ggtcacttcg atggtttgcc cggataaacg gaactggaaa aactgctgct ggtgttttgc
     2221 ttccgtcagc gctggatgcg gcgtgcggtc ggcaaagacc agaccgttca tacagaactg
     2281 gcgatcgttc ggcgtatcgc caaaatcacc gccgtaagcc gaccacgggt tgccgttttc
     2341 atcatattta atcagcgact gatccaccca gtcccagacg aagccgccct gtaaacgggg
     2401 atactgacga aacgcctgcc agtatttagc gaaaccgcca agactgttac ccatcgcgtg
     2461 ggcgtattcg caaaggatca gcgggcgcgt ctctccaggt agcgaaagcc attttttgat
     2521 ggaccatttc ggcacagccg ggaagggctg gtcttcatcc acgcgcgcgt acatcgggca
     2581 aataatatcg gtggccgtgg tgtcggctcc gccgccttca tactgcaccg ggcgggaagg
     2641 atcgacagat ttgatccagc gatacagcgc gtcgtgatta gcgccgtggc ctgattcatt
     2701 ccccagcgac cagatgatca cactcgggtg attacgatcg cgctgcacca ttcgcgttac
     2761 gcgttcgctc atcgccggta gccagcgcgg atcatcggtc agacgattca ttggcaccat
     2821 gccgtgggtt tcaatattgg cttcatccac cacatacagg ccgtagcggt cgcacagcgt
     2881 gtaccacagc ggatggttcg gataatgcga acagcgcacg gcgttaaagt tgttctgctt
     2941 catcagcagg atatcctgca ccatcgtctg ctcatccatg acctgaccat gcagaggatg
     3001 atgctcgtga cggttaacgc ctcgaatcag caacggcttg ccgttcagca gcagcagacc
     3061 attttcaatc cgcacctcgc ggaaaccgac atcgcaggct tctgcttcaa tcagcgtgcc
     3121 gtcggcggtg tgcagttcaa ccaccgcacg atagagattc gggatttcgg cgctccacag
     3181 tttcgggttt tcgacgttca gacgtagtgt gacgcgatcg gcataaccac cacgctcatc
     3241 gataatttca ccgccgaaag gcgcggtgcc gctggcgacc tgcgtttcac cctgccataa
     3301 agaaactgtt acccgtaggt agtcacgcaa ctcgccgcac atctgaactt cagcctccag
     3361 tacagcgcgg ctgaaatcat cattaaagcg agtggcaaca tggaaatcgc tgatttgtgt
     3421 agtcggttta tgcagcaacg agacgtcacg gaaaatgccg ctcatccgcc acatatcctg
     3481 atcttccaga taactgccgt cactccaacg cagcaccatc accgcgaggc ggttttctcc
     3541 ggcgcgtaaa aatgcgctca ggtcaaattc agacggcaaa cgactgtcct ggccgtaacc
     3601 gacccagcgc ccgttgcacc acagatgaaa cgccgagtta acgccatcaa aaataattcg
     3661 cgtctggcct tcctgtagcc agctttcatc aacattaaat gtgagcgagt aacaacccgt
     3721 cggattctcc gtgggaacaa acggcggatt gaccgtaatg ggataggtta cgttggtgta
     3781 gatgggcgca tcgtaaccgt gcatctgcca gtttgagggg acgacgacag tatcggcctc
     3841 aggaagatcg cactccagcc agctttccgg caccgcttct ggtgccggaa accaggcaaa
     3901 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     3961 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     4021 agggttttcc cagtcacgac gttgtaaaac gacgggatcg cgcttgagca gctccttgct
     4081 ggtgtccaga ccaatgcctc ccagaccggc aacgaaaatc acgttcttgt tggtcaaagt
     4141 aaacgacatg gtgacttctt ttttgcttta gcaggctctt tcgatccccg ggaattgcgg
     4201 ccgccgacta gtgagctcgt cgacccggga attccggacc ggtacctgca ggcgtacctt
     4261 ctatagtgtc acctaaatag ctttttgcaa aagcctaggc tagagtccgg aggctggatc
     4321 ggtcccggtg tcttctatgg aggtcaaaac agcgtggatg gcgtctccag gcgatctgac
     4381 ggttcactaa acgagctctg cttatataga cctcccaccg tacacgccta ccgcccattt
     4441 gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa
     4501 acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct
     4561 atccacgccc attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata
     4621 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     4681 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     4741 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     4801 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     4861 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gacctgcacg atgctgtttc
     4921 ctgtgtgaaa ttgttatccg ctcacaattc cacacattat acgagccgga agctataaag
     4981 tgtaaagcct ggggtgccta atgagtgaaa gggcctcgta tcacgcctat ttttataggt
     5041 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     5101 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     5161 ataaccctga taaatgcttc aataatattg aaaaacgcgc gaattgcaag ctctgcatta
     5221 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5281 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5341 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5401 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5461 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5521 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5581 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5641 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5701 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5761 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5821 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5881 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5941 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6001 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6061 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgccataact
     6121 tcgtatagca tacattatac gaagttatgg catgagatta tcaaaaagga tcttcaccta
     6181 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6241 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6301 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6361 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6421 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6481 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6541 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6601 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6661 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6721 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6781 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6841 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6901 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6961 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7021 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7081 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7141 ttatcagggt tattgtctca tgccaggggt gggcacacat atttgatacc agcgatccct
     7201 acacagcaca taattcaatg cgacttccct ctatcgcaca tcttagacct ttattctccc
     7261 tccagcacac atcgaagctg ccgagcaagc cgttctcacc agtccaagac ctggcatgag
     7321 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7381 ccgaaaagtg ccacctgaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt
     7441 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     7501 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     7561 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     7621 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     7681 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     7741 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     7801 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtc
//
