LOCUS       40924_14440        8930 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 8930)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8930)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8930
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    81..102
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        117..147
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    155..171
                     /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     179..195
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        216..234
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1018..1722
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1734..1754
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             1764..2105
                     /codon_start=1
                     /label=GAL4 activation domain
                     /note="activation domain of the GAL4 transcriptional
                     activator"
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     protein_bind    2121..2245
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2392..2494
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             2495..3172
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     CDS             3495..3797
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(3841..3965)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      4165..4352
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     promoter        complement(4505..4523)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4530..4546)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4687..5142
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(5248..5919)
                     /codon_start=1
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(5920..6201)
                     /label=TRP1 promoter
     misc_feature    complement(6459..6962)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     promoter        6999..7103
                     /label=AmpR promoter
     CDS             7104..7961
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      8135..8723
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
       61 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
      121 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
      181 ggaaacagct atgaccatga ttacgccaag ctcgaaatta accctcacta aagggaacaa
      241 aagctggtac cgggcccccc ctcgagatcc gggatcgaag aaatgatggt aaatgaaata
      301 ggaaatcaag gagcatgaag gcaaaagaca aatataaggg tcgaacgaaa aataaagtga
      361 aaagtgttga tatgatgtat ttggctttgc ggcgccgaaa aaacgagttt acgcaattgc
      421 acaatcatgc tgactctgtg gcggacccgc gctcttgccg gcccggcgat aacgctgggc
      481 gtgaggctgt gcccggcgga gttttttgcg cctgcatttt ccaaggttta ccctgcgcta
      541 aggggcgaga ttggagaagc aataagaatg ccggttgggg ttgcgatgat gacgaccacg
      601 acaactggtg tcattattta agttgccgaa agaacctgag tgcatttgca acatgagtat
      661 actagaagaa tgagccaaga cttgcgagac gcgagtttgc cggtggtgcg aacaatagag
      721 cgaccatgac cttgaaggtg agacgcgcat aaccgctaga gtactttgaa gaggaaacag
      781 caatagggtt gctaccagta taaatagaca ggtacataca acactggaaa tggttgtctg
      841 tttgagtacg ctttcaattc atttgggtgt gcactttatt atgttacaat atggaaggga
      901 actttacact tctcctatgc acatatatta attaaagtcc aatgctagta gagaaggggg
      961 gtaacacccc tccgcgctct tttccgattt ttttctaaac cgtggaatat ttcggatatc
     1021 cttttgttgt ttccgggtgt acaatatgga cttcctcttt tctggcaacc aaacccatac
     1081 atcgggattc ctataatacc ttcgttggtc tccctaacat gtaggtggcg gaggggagat
     1141 atacaataga acagatacca gacaagacat aatgggctaa acaagactac accaattaca
     1201 ctgcctcatt gatggtggta cataacgaac taatactgta gccctagact tgatagccat
     1261 catcatatcg aagtttcact accctttttc catttgccat ctattgaagt aataataggc
     1321 gcatgcaact tcttttcttt ttttttcttt tctctctccc ccgttgttgt ctcaccatat
     1381 ccgcaatgac aaaaaaatga tggaagacac taaaggaaaa aattaacgac aaagacagca
     1441 ccaacagatg tcgttgttcc agagctgatg aggggtatct cgaagcacac gaaacttttt
     1501 ccttccttca ttcacgcaca ctactctcta atgagcaacg gtatacggcc ttccttccag
     1561 ttacttgaat ttgaaataaa aaaaagtttg ctgtcttgct atcaagtata aatagacctg
     1621 caattattaa tcttttgttt cctcgtcatt gttctcgttc cctttcttcc ttgtttcttt
     1681 ttctgcacaa tatttcaagc tataccaagc atacaatcaa ctccaagctt atgcccaaga
     1741 agaagcggaa ggtctcgagc ggcgccaatt ttaatcaaag tgggaatatt gctgatagct
     1801 cattgtcctt cactttcact aacagtagca acggtccgaa cctcataaca actcaaacaa
     1861 attctcaagc gctttcacaa ccaattgcct cctctaacgt tcatgataac ttcatgaata
     1921 atgaaatcac ggctagtaaa attgatgatg gtaataattc aaaaccactg tcacctggtt
     1981 ggacggacca aactgcgtat aacgcgtttg gaatcactac agggatgttt aataccacta
     2041 caatggatga tgtatataac tatctattcg atgatgaaga taccccacca aacccaaaaa
     2101 aagagggtgg gtcgaatcaa acaagtttgt acaaaaaagc tgaacgagaa acgtaaaatg
     2161 atataaatat caatatatta aattagattt tgcataaaaa acagactaca taatactgta
     2221 aaacacaaca tatccagtca ctatggcggc cgctaagttg gcagcatcac ccgacgcact
     2281 ttgcgccgaa taaatacctg tgacggaaga tcacttcgca gaataaataa atcctggtgt
     2341 ccctgttgat accgggaagc cctgggccaa cttttggcga aaatgagacg ttgatcggca
     2401 cgtaagaggt tccaactttc accataatga aataagatca ctaccgggcg tattttttga
     2461 gtcatcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac
     2521 caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc
     2581 tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa
     2641 gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc
     2701 tcatccggaa ttccgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca
     2761 cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggagtgaata
     2821 ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga
     2881 aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct cagccaatcc
     2941 ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc
     3001 cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat
     3061 tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta atgaattaca
     3121 acagtactgc gatgagtggc aggcggggcg taatctagag gatccggctt actaaaagcc
     3181 agataacagt atgcgtattt gcgcgctgat ttttgcggta taagaatata tactgatatg
     3241 tatacccgaa gtatgtcaaa aagaggtatg ctatgaagca gcgtattaca gtgacagttg
     3301 acagcgacag ctatcagttg ctcaaggcat atatgatgtc aatatctccg gtctggtaag
     3361 cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa cgctggaaag cggaaaatca
     3421 ggaagggatg gctgaggtcg cccggtttat tgaaatgaac ggctcttttg ctgacgagaa
     3481 caggggctgg tgaaatgcag tttaaggttt acacctataa aagagagagc cgttatcgtc
     3541 tgtttgtgga tgtacagagt gatattattg acacgcccgg gcgacggatg gtgatccccc
     3601 tggccagtgc acgtctgctg tcagataaag tcccccgtga actttacccg gtggtgcata
     3661 tcggggatga aagctggcgc atgatgacca ccgatatggc cagtgtgccg gtctccgtta
     3721 tcggggaaga agtggctgat ctcagccacc gcgaaaatga catcaaaaac gccattaacc
     3781 tgatgttctg gggaatataa atgtcaggct cccttataca cagccagtct gcaggtcgac
     3841 catagtgact ggatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc
     3901 taatttaata tattgatatt tatatcattt tacgtttctc gttcagcttt cttgtacaaa
     3961 gtggtttgat ggccgctaag taagtaagac gtcgagctct aagtaagtaa cggccgccac
     4021 cgcggtggag ctttggactt cttcgccaga ggtttggtca agtctccaat caaggttgtc
     4081 ggcttgtcta ccttgccaga aatttacgaa aagatggaaa agggtcaaat cgttggtaga
     4141 tacgttgttg acacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa
     4201 taagttataa aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga
     4261 aaattcttat tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagcat
     4321 gaggtcgctc ttattgacca cacctctacc ggcatgccga gcaaatgcct gcaaatcgct
     4381 ccccatttca cccaattgta gatatgctaa ctccagcaat gagttgatga atctcggtgt
     4441 gtattttatg tcctcagagg acaacacctg ttgtaatcgt tcttccacac ggatcccaat
     4501 tcgccctata gtgagtcgta ttacaattca ctggccgtcg ttttacaacg tcgtgactgg
     4561 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     4621 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     4681 gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4741 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     4801 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4861 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     4921 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4981 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     5041 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     5101 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct
     5161 ccttacgcat ctgtgcggta tttcacaccg catagatcgg caagtgcaca aacaatactt
     5221 aaataaatac tactcagtaa taacctattt cttagcattt ttgacgaaat ttgctatttt
     5281 gttagagtct tttacaccat ttgtctccac acctccgctt acatcaacac caataacgcc
     5341 atttaatcta agcgcatcac caacattttc tggcgtcagt ccaccagcta acataaaatg
     5401 taagctttcg gggctctctt gccttccaac ccagtcagaa atcgagttcc aatccaaaag
     5461 ttcacctgtc ccacctgctt ctgaatcaaa caagggaata aacgaatgag gtttctgtga
     5521 agctgcactg agtagtatgt tgcagtcttt tggaaatacg agtcttttaa taactggcaa
     5581 accgaggaac tcttggtatt cttgccacga ctcatctcca tgcagttgga cgatatcaat
     5641 gccgtaatca ttgaccagag ccaaaacatc ctccttaggt tgattacgaa acacgccaac
     5701 caagtatttc ggagtgcctg aactattttt atatgctttt acaagacttg aaattttcct
     5761 tgcaataacc gggtcaattg ttctctttct attgggcaca catataatac ccagcaagtc
     5821 agcatcggaa tctagagcac attctgcggc ctctgtgctc tgcaagccgc aaactttcac
     5881 caatggacca gaactacctg tgaaattaat aacagacata ctccaagctg cctttgtgtg
     5941 cttaatcacg tatactcacg tgctcaatag tcaccaatgc cctccctctt ggccctctcc
     6001 ttttcttttt tcgaccgaat taattcttaa tcggcaaaaa aagaaaagct ccggatcaag
     6061 attgtacgta aggtgacaag ctatttttca ataaagaata tcttccacta ctgccatctg
     6121 gcgtcataac tgcaaagtac acatatatta cgatgctgtt ctattaaatg cttcctatat
     6181 tatatatata gtaatgtcgt gatctatggt gcactctcag tacaatctgc tctgatgccg
     6241 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     6301 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     6361 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     6421 tataggttaa tgtcatgata ataatggttt cttagacgga tcgcttgcct gtaacttaca
     6481 cgcgcctcgt atcttttaat gatggaataa tttgggaatt tactctgtgt ttatttattt
     6541 ttatgttttg tatttggatt ttagaaagta aataaagaag gtagaagagt tacggaatga
     6601 agaaaaaaaa ataaacaaag gtttaaaaaa tttcaacaaa aagcgtactt tacatatata
     6661 tttattagac aagaaaagca gattaaatag atatacattc gattaacgat aagtaaaatg
     6721 taaaatcaca ggattttcgt gtgtggtctt ctacacagac aagatgaaac aattcggcat
     6781 taatacctga gagcaggaag agcaagataa aaggtagtat ttgttggcga tccccctaga
     6841 gtcttttaca tcttcggaaa acaaaaacta ttttttcttt aatttctttt tttactttct
     6901 atttttaatt tatatattta tattaaaaaa tttaaattat aattattttt atagcacgtg
     6961 atgaaaagga cccaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     7021 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     7081 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     7141 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     7201 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     7261 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     7321 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     7381 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     7441 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     7501 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     7561 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     7621 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     7681 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     7741 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     7801 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     7861 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     7921 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     7981 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     8041 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     8101 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     8161 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     8221 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     8281 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     8341 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     8401 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     8461 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     8521 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     8581 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     8641 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     8701 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     8761 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     8821 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     8881 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
//
