LOCUS       62056_8140        6871 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 6871)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        494..598
                     /label=AmpR promoter
     CDS             599..1456
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1630..2218
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2404..2546)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2955..3128)
                     /codon_start=1
                     /label=lacZ-alpha
                     /note="LacZ-alpha fragment of beta-galactosidase"
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGE"
     protein_bind    complement(3148..3164)
                     /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(3172..3202)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3217..3238)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3254..4333)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(4334..4411)
                     /label=lacI promoter
     promoter        4529..4557
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    4565..4581
                     /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)."
     protein_bind    complement(4733..4766)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4781..5226
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             5227..6645
                     /codon_start=1
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt
      481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      541 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
      601 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
      661 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
      721 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
      781 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
      841 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
      901 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
      961 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     1021 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     1081 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     1141 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     1201 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     1261 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     1321 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     1381 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     1441 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     1501 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1561 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1621 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1681 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1741 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     1801 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1861 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1921 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1981 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2041 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2101 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2161 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2221 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2281 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     2341 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     2401 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatatgg
     2461 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     2521 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     2581 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     2641 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagaac gccatcaaaa
     2701 ataattcgcg tctggccttc ctgtagccag ctttcatcaa cattaaatgt gagcgagtaa
     2761 caacccgtcg gattctccgt gggaacaaac ggcggattga ccgtaatggg ataggttacg
     2821 ttggtgtaga tgggcgcatc gtaaccgtgc atctgccagt ttgaggggac gacgacagta
     2881 tcggcctcag gaagatcgca ctccagccag ctttccggca ccgcttctgg tgccggaaac
     2941 caggcaaagc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     3001 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     3061 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga atccgtaatc
     3121 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     3181 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     3241 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3301 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
     3361 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
     3421 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
     3481 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac
     3541 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
     3601 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
     3661 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
     3721 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
     3781 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
     3841 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
     3901 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
     3961 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
     4021 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
     4081 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
     4141 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
     4201 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
     4261 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
     4321 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     4381 cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac
     4441 tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg
     4501 aatggtgcat gctcacgagg tcctcgagtt gacaattaat catcggctcg tataatgtgt
     4561 ggaattgtga gcggataaca atttcacaca ggaaacagga tcagcttact ccccatcccc
     4621 ctgggatcca tcgatgcttt tgacaattaa tcatcggctc gtataatgtg tggaattgtg
     4681 agcggataac aaaggaggtc atatgattat gagttctcct cctgaaagat ccataacttc
     4741 gtatagcata cattatacga agttatgcgg ccgcgacgtc cacatatacc tgccgttcac
     4801 tattatttag tgaaatgaga tattatgata ttttctgaat tgtgattaaa aaggcaactt
     4861 tatgcccatg caacagaaac tataaaaaat acagagaatg aaaagaaaca gatagatttt
     4921 ttagttcttt aggcccgtag tctgcaaatc cttttatgat tttctatcaa acaaaagagg
     4981 aaaatagacc agttgcaatc caaacgagag tctaatagaa tgaggtcgaa aagtaaatcg
     5041 cgcgggtttg ttactgataa agcaggcaag acctaaaatg tgtaaagggc aaagtgtata
     5101 ctttggcgtc accccttaca tattttaggt ctttttttat tgtgcgtaac taacttgcca
     5161 tcttcaaaca ggagggctgg aagaagcaga ccgctaacac agtacataaa aaaggagaca
     5221 tgaacgatga acatcaaaaa gtttgcaaaa caagcaacag tattaacctt tactaccgca
     5281 ctgctggcag gaggcgcaac tcaagcgttt gcgaaagaaa cgaaccaaaa gccatataag
     5341 gaaacatacg gcatttccca tattacacgc catgatatgc tgcaaatccc tgaacagcaa
     5401 aaaaatgaaa aatatcaagt tcctgagttc gattcgtcca caattaaaaa tatctcttct
     5461 gcaaaaggcc tggacgtttg ggacagctgg ccattacaaa acgctgacgg cactgtcgca
     5521 aactatcacg gctaccacat cgtctttgca ttagccggag atcctaaaaa tgcggatgac
     5581 acatcgattt acatgttcta tcaaaaagtc ggcgaaactt ctattgacag ctggaaaaac
     5641 gctggccgcg tctttaaaga cagcgacaaa ttcgatgcaa atgattctat cctaaaagac
     5701 caaacacaag aatggtcagg ttcagccaca tttacatctg acggaaaaat ccgtttattc
     5761 tacactgatt tctccggtaa acattacggc aaacaaacac tgacaactgc acaagttaac
     5821 gtatcagcat cagacagctc tttgaacatc aacggtgtag aggattataa atcaatcttt
     5881 gacggtgacg gaaaaacgta tcaaaatgta cagcagttca tcgatgaagg caactacagc
     5941 tcaggcgaca accatacgct gagagatcct cactacgtag aagataaagg ccacaaatac
     6001 ttagtatttg aagcaaacac tggaactgaa gatggctacc aaggcgaaga atctttattt
     6061 aacaaagcat actatggcaa aagcacatca ttcttccgtc aagaaagtca aaaacttctg
     6121 caaagcgata aaaaacgcac ggctgagtta gcaaacggcg ctctcggtat gattgagcta
     6181 aacgatgatt acacactgaa aaaagtgatg aaaccgctga ttgcatctaa cacagtaaca
     6241 gatgaaattg aacgcgcgaa cgtctttaaa atgaacggca aatggtacct gttcactgac
     6301 tcccgcggat caaaaatgac gattgacggc attacgtcta acgatattta catgcttggt
     6361 tatgtttcta attctttaac tggcccatac aagccgctga acaaaactgg ccttgtgtta
     6421 aaaatggatc ttgatcctaa cgatgtaacc tttacttact cacacttcgc tgtacctcaa
     6481 gcgaaaggaa acaatgtcgt gattacaagc tatatgacaa acagaggatt ctacgcagac
     6541 aaacaatcaa cgtttgcgcc gagcttcctg ctgaacatca aaggcaagaa aacatctgtt
     6601 gtcaaagaca gcatccttga acaaggacaa ttaacagtta acaaataaaa acgcaaaaga
     6661 aaatgccgat attgacgtca ggtggcactt ttcgaggaga tcatgcacat gatgacgaag
     6721 cttatcgatg ataagctgtc aaacatgagc agatctgagc ccgcctaatg agcgggcttt
     6781 tagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     6841 ttcttaggtg ctcgtcaaag caactccgga t
//
