LOCUS       40924_16935        9226 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 9226)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 9226)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9226
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        475..1653
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     promoter        1670..1688
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1782..4826
                     /codon_start=1
                     /label=lacZ
                     /note="beta-galactosidase"
                     /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS
                     LNGEWRFAWFPVPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNPP
                     FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF
                     DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR
                     FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV
                     TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP
                     LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL
                     YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA
                     NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS
                     LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP
                     WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL
                     FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ
                     PNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ
                     SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA
                     ALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP
                     ARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGLR
                     CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD
                     SWSPSVSAEFQLSAGRYHYQLVWCQK"
     CDS             4851..4880
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     CDS             4896..4913
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     polyA_signal    4942..5166
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5212..5640
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5654..5984
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             6051..6842
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    7021..7154
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7191..7207)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(7215..7231)
                     /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(7239..7269)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7284..7305)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7593..8181)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8355..9212)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(join(9213..9226,1..91))
                     /label=AmpR promoter
ORIGIN
        1 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
       61 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
      121 tcgacggatc gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga
      181 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      241 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      301 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      361 ttgattattg actaggcttt tgcaaaaagc tttgcaaaga tggataaagt tttaaacaga
      421 gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgcctc gtgaggctcc
      481 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
      541 gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
      601 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
      661 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt
      721 ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc
      781 acctggctgc agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt
      841 tcgaggcctt gcgcttaagg agccccttcg cctcgtgctt gagttgaggc ctggcctggg
      901 cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat
      961 aagtctctag ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat
     1021 agtcttgtaa atgcgggcca agatctgcac actggtattt cggtttttgg ggccgcgggc
     1081 ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg
     1141 ccaccgagaa tcggacgggg gtagtctcaa gctggccggc ctgctctggt gcctggcctc
     1201 gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg
     1261 tgagcggaaa gatggccgct tcccggccct gctgcaggga gctcaaaatg gaggacgcgg
     1321 cgctcgggag agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca
     1381 gccgtcgctt catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc
     1441 tcgagctttt ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt
     1501 ccccacactg agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc
     1561 ttggaatttg ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt
     1621 caaagttttt ttcttccatt tcaggtgtcg tgaggaatta gcttggtact aatacgactc
     1681 actataggga gacccaagct ggctagttaa gcttggtacc gagctcggat ccactagtcc
     1741 agtgtggtgg aattctgcag atcgaaacga tgatagatcc cgtcgtttta caacgtcgtg
     1801 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     1861 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     1921 atggcgaatg gcgctttgcc tggtttccgg taccagaagc ggtgccggaa agctggctgg
     1981 agtgcgatct tcctgaggcc gatactgtcg tcgtcccctc aaactggcag atgcacggtt
     2041 acgatgcgcc catctacacc aacgtaacct atcccattac ggtcaatccg ccgtttgttc
     2101 ccacggagaa tccgacgggt tgttactcgc tcacatttaa tgttgatgaa agctggctac
     2161 aggaaggcca gacgcgaatt atttttgatg gcgttaactc ggcgtttcat ctgtggtgca
     2221 acgggcgctg ggtcggttac ggccaggaca gtcgtttgcc gtctgaattt gacctgagcg
     2281 catttttacg cgccggagaa aaccgcctcg cggtgatggt gctgcgttgg agtgacggca
     2341 gttatctgga agatcaggat atgtggcgga tgagcggcat tttccgtgac gtctcgttgc
     2401 tgcataaacc gactacacaa atcagcgatt tccatgttgc cactcgcttt aatgatgatt
     2461 tcagccgcgc tgtactggag gctgaagttc agatgtgcgg cgagttgcgt gactacctac
     2521 gggtaacagt ttctttatgg cagggtgaaa cgcaggtcgc cagcggcacc gcgcctttcg
     2581 gcggtgaaat tatcgatgag cgtggtggtt atgccgatcg cgtcacacta cgtctgaacg
     2641 tcgaaaaccc gaaactgtgg agcgccgaaa tcccgaatct ctatcgtgcg gtggttgaac
     2701 tgcacaccgc cgacggcacg ctgattgaag cagaagcctg cgatgtcggt ttccgcgagg
     2761 tgcggattga aaatggtctg ctgctgctga acggcaagcc gttgctgatt cgaggcgtta
     2821 accgtcacga gcatcatcct ctgcatggtc aggtcatgga tgagcagacg atggtgcagg
     2881 atatcctgct gatgaagcag aacaacttta acgccgtgcg ctgttcgcat tatccgaacc
     2941 atccgctgtg gtacacgctg tgcgaccgct acggcctgta tgtggtggat gaagccaata
     3001 ttgaaaccca cggcatggtg ccaatgaatc gtctgaccga tgatccgcgc tggctaccgg
     3061 cgatgagcga acgcgtaacg cgaatggtgc agcgcgatcg taatcacccg agtgtgatca
     3121 tctggtcgct ggggaatgaa tcaggccacg gcgctaatca cgacgcgctg tatcgctgga
     3181 tcaaatctgt cgatccttcc cgcccggtgc agtatgaagg cggcggagcc gacaccacgg
     3241 ccaccgatat tatttgcccg atgtacgcgc gcgtggatga agaccagccc ttcccggctg
     3301 tgccgaaatg gtccatcaaa aaatggcttt cgctacctgg agagacgcgc ccgctgatcc
     3361 tttgcgaata cgcccacgcg atgggtaaca gtcttggcgg tttcgctaaa tactggcagg
     3421 cgtttcgtca gtatccccgt ttacagggcg gcttcgtctg ggactgggtg gatcagtcgc
     3481 tgattaaata tgatgaaaac ggcaacccgt ggtcggctta cggcggtgat tttggcgata
     3541 cgccgaacga tcgccagttc tgtatgaacg gtctggtctt tgccgaccgc acgccgcatc
     3601 cagcgctgac ggaagcaaaa caccagcagc agtttttcca gttccgttta tccgggcaaa
     3661 ccatcgaagt gaccagcgaa tacctgttcc gtcatagcga taacgagctc ctgcactgga
     3721 tggtggcgct ggatggtaag ccgctggcaa gcggtgaagt gcctctggat gtcgctccac
     3781 aaggtaaaca gttgattgaa ctgcctgaac taccgcagcc ggagagcgcc gggcaactct
     3841 ggctcacagt acgcgtagtg caaccgaacg cgaccgcatg gtcagaagcc gggcacatca
     3901 gcgcctggca gcagtggcgt ctggcggaaa acctcagtgt gacgctcccc gccgcgtccc
     3961 acgccatccc gcatctgacc accagcgaaa tggatttttg catcgagctg ggtaataagc
     4021 gttggcaatt taaccgccag tcaggctttc tttcacagat gtggattggc gataaaaaac
     4081 aactgctgac gccgctgcgc gatcagttca cccgtgcacc gctggataac gacattggcg
     4141 taagtgaagc gacccgcatt gaccctaacg cctgggtcga acgctggaag gcggcgggcc
     4201 attaccaggc cgaagcagcg ttgttgcagt gcacggcaga tacacttgct gatgcggtgc
     4261 tgattacgac cgctcacgcg tggcagcatc aggggaaaac cttatttatc agccggaaaa
     4321 cctaccggat tgatggtagt ggtcaaatgg cgattaccgt tgatgttgaa gtggcgagcg
     4381 atacaccgca tccggcgcgg attggcctga actgccagct ggcgcaggta gcagagcggg
     4441 taaactggct cggattaggg ccgcaagaaa actatcccga ccgccttact gccgcctgtt
     4501 ttgaccgctg ggatctgcca ttgtcagaca tgtatacccc gtacgtcttc ccgagcgaaa
     4561 acggtctgcg ctgcgggacg cgcgaattga attatggccc acaccagtgg cgcggcgact
     4621 tccagttcaa catcagccgc tacagtcaac agcaactgat ggaaaccagc catcgccatc
     4681 tgctgcacgc ggaagaaggc acatggctga atatcgacgg tttccatatg gggattggtg
     4741 gcgacgactc ctggagcccg tcagtatcgg cggaattcca gctgagcgcc ggtcgctacc
     4801 attaccagtt ggtctggtgt caaaaagcgg ccgctcgagg tcacccattc gaacaaaaac
     4861 tcatctcaga agaggatctg aatatgcata ccggtcatca tcaccatcac cattgagttt
     4921 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     4981 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     5041 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     5101 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     5161 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     5221 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     5281 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     5341 gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac
     5401 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     5461 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     5521 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     5581 tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     5641 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc caggcaggca
     5701 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     5761 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     5821 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     5881 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     5941 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     6001 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     6061 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     6121 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     6181 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     6241 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     6301 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     6361 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     6421 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     6481 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     6541 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     6601 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     6661 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     6721 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     6781 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     6841 tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca
     6901 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     6961 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     7021 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     7081 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     7141 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     7201 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     7261 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     7321 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     7381 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     7441 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     7501 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     7561 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     7621 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     7681 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     7741 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt
     7801 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     7861 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     7921 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     7981 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     8041 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     8101 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     8161 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     8221 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     8281 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     8341 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     8401 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     8461 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     8521 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     8581 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     8641 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     8701 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     8761 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     8821 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     8881 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     8941 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     9001 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     9061 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     9121 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     9181 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttc
//
