LOCUS       40924_14070        6846 bp DNA     circular SYN 13-JAN-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6846)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6846)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6846
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..80
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             81..1160
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    1176..1197
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        1406..1434
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    1442..1458
                     /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)."
     CDS             1576..2394
                     /codon_start=1
                     /label=Sce VMA intein 5' region
                     /note="5' region of the intein from the yeast Vma1 subunit
                     of the vacuolar ATPase (Chong et al., 1998)"
                     /translation="CFAKGTNVLMADGSIECIENIEVGNKVMGKDGRPREVIKLPRGRE
                     TMYSVVQKSQHRAHKSDSSREVPELLKFTCNATHELVVRTPRSVRRLSRTIKGVEYFEV
                     ITFEMGQKKAPDGRIVELVKEVSKSYPISEGPERANELVESYRKASNKAYFEWTIEARD
                     LSLLGSHVRKATYQTYAPILYENDHFFDYMQKSKFHLTIEGPKVLAYLLGLWIGDGLSD
                     RATFSVDSRDTSLMERVTEYAEKLNLCAEYKDRKEPQVAKTVNLYSKVVRG"
     CDS             2398..2931
                     /codon_start=1
                     /label=Sce VMA intein 3' region
                     /note="modified 3' region of the intein from the yeast Vma1
                     subunit of the vacuolar ATPase (Chong et al., 1998)"
                     /translation="GIRNNLNTENPLWDAIVGLGFLKDGVKNIPSFLSTDNIGTRETFL
                     AGLIDSDGYVTDEHGIKATIKTIHTSVRDGLVSLARSLGLVVSVNAEPAKVDMNVTKHK
                     ISYAIYMSGGDVLLNVLSKCAGSKKFRPAPAAAFARECRGFYFELQELKEDDYYGITLS
                     DDSDHQFLLGSQVVV"
     CDS             2971..3126
                     /codon_start=1
                     /label=CBD
                     /note="chitin binding domain from chitinase A1 (Watanabe et
                     al., 1994)"
                     /translation="TTNPGVSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSLAGWEPSNV
                     PALWQLQ"
     terminator      3360..3446
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3538..3565
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        3585..3676
                     /label=AmpR promoter
     CDS             3677..4534
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      complement(4579..5092)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      5203..5791
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6163..6351)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ccgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
       61 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
      121 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
      181 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
      241 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
      301 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
      361 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
      421 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
      481 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
      541 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
      601 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
      661 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
      721 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
      781 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
      841 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg
      901 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
      961 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     1021 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
     1081 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     1141 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     1201 gcacaattct catgtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg
     1261 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg
     1321 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt
     1381 ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga
     1441 attgtgagcg gataacaatt tcacacagga aacagccagt ccgtttaggt gttttcacga
     1501 gcacttcacc aacaaggacc atagcatatg gctagctcgc gagtcgactc tagagaattc
     1561 ctcgagggct cttcctgctt tgccaagggt accaatgttt taatggcgga tgggtctatt
     1621 gaatgtattg aaaacattga ggttggtaat aaggtcatgg gtaaagatgg cagacctcgt
     1681 gaggtaatta aattgcccag aggaagagaa actatgtaca gcgtcgtgca gaaaagtcag
     1741 cacagagccc acaaaagtga ctcaagtcgt gaagtgccag aattactcaa gtttacgtgt
     1801 aatgcgaccc atgagttggt tgttagaaca cctcgtagtg tccgccgttt gtctcgtacc
     1861 attaagggtg tcgaatattt tgaagttatt acttttgaga tgggccaaaa gaaagccccc
     1921 gacggtagaa ttgttgagct tgtcaaggaa gtttcaaaga gctacccaat atctgagggg
     1981 cctgagagag ccaacgaatt agtagaatcc tatagaaagg cttcaaataa agcttatttt
     2041 gagtggacta ttgaggccag agatctttct ctgttgggtt cccatgttcg taaagctacc
     2101 taccagactt acgctccaat tctttatgag aatgaccact ttttcgacta catgcaaaaa
     2161 agtaagtttc atctcaccat tgaaggtcca aaagtacttg cttatttact tggtttatgg
     2221 attggtgatg gattgtctga cagggcaact ttttcggttg attccagaga tacttctttg
     2281 atggaacgtg ttactgaata tgctgaaaag ttgaatttgt gcgccgagta taaggacaga
     2341 aaagaaccac aagttgccaa aactgttaat ttgtactcta aagttgtcag aggtaatggt
     2401 attcgcaata atcttaatac tgagaatcca ttatgggacg ctattgttgg cttaggattc
     2461 ttgaaggacg gtgtcaaaaa tattccttct ttcttgtcta cggacaatat cggtactcgt
     2521 gaaacatttc ttgctggtct aattgattct gatggctatg ttactgatga gcatggtatt
     2581 aaagcaacaa taaagacaat tcatacttct gtcagagatg gtttggtttc ccttgctcgt
     2641 tctttaggct tagtagtctc ggttaacgca gaacctgcta aggttgacat gaatgtcacc
     2701 aaacataaaa ttagttatgc tatttatatg tctggtggag atgttttgct taacgttctt
     2761 tcgaagtgtg ccggctctaa aaaattcagg cctgctcccg ccgctgcttt tgcacgtgag
     2821 tgccgcggat tttatttcga gttacaagaa ttgaaggaag acgattatta tgggattact
     2881 ttatctgatg attctgatca tcagtttttg cttggatccc aggttgtcgt ccatgcatgc
     2941 ggtggcctga ccggtctgaa ctcaggcctc acgacaaatc ctggtgtatc cgcttggcag
     3001 gtcaacacag cttatactgc gggacaattg gtcacatata acggcaagac gtataaatgt
     3061 ttgcagcccc acacctcctt ggcaggatgg gaaccatcca acgttcctgc cttgtggcag
     3121 cttcaatgac tgcagcctga atggcgaatg gcagcttggc tgttttggcg gatgagataa
     3181 gattttcagc ctgatacaga ttaaatcaga acgcagaagc ggtctgataa aacagaattt
     3241 gcctggcggc agtagcgcgg tggtcccacc tgaccccatg ccgaactcag aagtgaaacg
     3301 ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc
     3361 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     3421 tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac
     3481 ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga
     3541 aggccatcct gacggatggc ctttttgcgt ttctacaaac tctttttgtt tatttttcta
     3601 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3661 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3721 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3781 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3841 tgagagtttt cgccccgaag aacgttctcc aatgatgagc acttttaaag ttctgctatg
     3901 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     3961 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     4021 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4081 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4141 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4201 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgtaaactat taactggcga
     4261 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4321 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4381 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4441 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4501 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     4561 tatactttag attgatttac cccggttgat aatcagaaaa gccccaaaaa caggaagatt
     4621 gtataagcaa atatttaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt
     4681 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     4741 aaagaatagc ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     4801 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     4861 cgtgaaccat cacccaaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     4921 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     4981 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     5041 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtaaaaggat
     5101 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5161 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5221 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5281 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5341 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5401 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5461 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5521 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5581 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5641 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5701 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5761 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5821 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5881 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     5941 gcgcagcgag tcagtgagcg aggaagctat ggtgcactct cagtacaatc tgctctgatg
     6001 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     6061 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     6121 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     6181 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgca gcgattcaca
     6241 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     6301 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca cttgatgcct
     6361 ccgtgtaagg gggaatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg
     6421 ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa
     6481 caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc
     6541 ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc
     6601 cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa
     6661 ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac
     6721 gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc
     6781 cgggtcctca acgacaggag cacgatcatg cgcacccgtg gccaggaccc aacgctgccc
     6841 gaaatt
//
