LOCUS       Exported                7650 bp DNA     circular SYN 06-JUN-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pT7-YME1L1(human)-6¡ÁHis
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7650)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7650
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    775..914
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(1016..1207)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2016..3098)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(3099..3176)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        3485..3503
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3504..3528
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     RBS             3543..3565
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3584..3601
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3611..3628
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             3632..3664
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             3674..5992
                     /codon_start=1
                     /label=YME1L1
                     /translation="MFSLSSTVQPQVTVPLSHLINAFHTPKNTSVSLSGVSVSQNQHRD
                     VVPEHEAPSSECMFSDFLTKLNIVSIGKGKIFEGYRSMFMEPAKRMKKSLDTTDNWHIR
                     PEPFSLSIPPSLNLRDLGLSELKIGQIDQLVENLLPGFCKGKNISSHWHTSHVSAQSFF
                     ENKYGNLDIFSTLRSSCLYRHHSRALQSICSDLQYWPVFIQSRGFKTLKSRTRRLQSTS
                     ERLAETQNIAPSFVKGFLLRDRGSDVESLDKLMKTKNIPEAHQDAFKTGFAEGFLKAQA
                     LTQKTNDSLRRTRLILFVLLLFGIYGLLKNPFLSVRFRTTTGLDSAVDPVQMKNVTFEH
                     VKGVEEAKQELQEVVEFLKNPQKFTILGGKLPKGILLVGPPGTGKTLLARAVAGEADVP
                     FYYASGSEFDEMFVGVGASRIRNLFREAKANAPCVIFIDELDSVGGKRIESPMHPYSRQ
                     TINQLLAEMDGFKPNEGVIIIGATNFPEALDNALIRPGRFDMQVTVPRPDVKGRTEILK
                     WYLNKIKFDQSVDPEIIARGTVGFSGAELENLVNQAALKAAVDGKEMVTMKELEFSKDK
                     ILMGPERRSVEIDNKNKTITAYHESGHAIIAYYTKDAMPINKATIMPRGPTLGHVSLLP
                     ENDRWNETRAQLLAQMDVSMGGRVAEELIFGTDHITTGASSDFDNATKIAKRMVTKFGM
                     SEKLGVMTYSDTGKLSPETQSAIEQEIRILLRDSYERAKHILKTHAKEHKNLAEALLTY
                     ETLDAKEIQIVLEGKKLEVR"
     CDS             5999..6016
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      6083..6130
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      6167..6622
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(6714..7529)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
      781 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt
      841 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
      901 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
      961 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     1021 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     1081 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     1141 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     1201 tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga
     1261 aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac
     1321 gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg
     1381 gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc
     1441 ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt
     1501 acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc
     1561 agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac
     1621 cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggggccg
     1681 ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga
     1741 aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg atcatcgtcg
     1801 cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc acctgtccta
     1861 cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg ccccgcgccc
     1921 accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct
     1981 aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     2041 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     2101 ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc
     2161 accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga
     2221 aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg
     2281 tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt
     2341 gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc
     2401 agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct
     2461 atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc
     2521 gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga
     2581 tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc
     2641 tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg
     2701 gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga
     2761 ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg
     2821 ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc
     2881 agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt
     2941 gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt
     3001 ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg
     3061 gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc
     3121 tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg
     3181 atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag
     3241 gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc
     3301 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
     3361 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
     3421 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg
     3481 aaattaatac gactcactat aggggaattg tgagcggata acaattcccc tctagaaata
     3541 attttgttta actttaagaa ggagatatac catgggcagc agccatcatc atcatcatca
     3601 cagcagcggc ctggtgccgc gcggcagcca tatggctagc atgactggtg gacagcaaat
     3661 gggtcgcgga tccatgtttt ccttgtcgag cacggtgcaa ccccaggtta cagttcctct
     3721 gagtcatctc atcaatgcct tccatacacc aaaaaacact tctgtttctc tcagtggagt
     3781 gtcagtttct caaaaccagc atcgagatgt agttcctgag catgaggctc ccagcagtga
     3841 gtgtatgttc agtgacttcc tgacgaagct taacattgtt tcaattggca aaggaaaaat
     3901 attcgaaggg tacagatcca tgttcatgga gccagcaaaa aggatgaaga agagcttgga
     3961 cacaaccgat aactggcaca tccgtccaga acccttctcc ctctcaatcc ctccttcact
     4021 taacttaagg gaccttggat tatctgaact aaaaattgga cagattgatc agctggtaga
     4081 aaatctactt cctggatttt gtaaaggcaa aaacatttct tcccattggc atacatccca
     4141 tgtctctgca caatccttct ttgaaaataa atatggtaac ttagatatat ttagtacatt
     4201 acgttcctct tgcttgtatc gacatcattc aagagctctt caaagcattt gttcagatct
     4261 tcagtactgg ccagttttca tacagtctcg gggttttaaa actttgaaat caaggacacg
     4321 acgtctccag tctacctccg agagattagc tgaaacacag aatatagcgc catcattcgt
     4381 gaaggggttt cttttgcggg acagaggatc agatgttgag agtttggaca aactcatgaa
     4441 aaccaaaaat atacctgaag ctcaccaaga tgcatttaaa actggttttg cggaaggttt
     4501 tctgaaagct caagcactca cacaaaaaac caatgattcc ctaaggcgaa cccgtctgat
     4561 tctcttcgtt ctgctgctat tcggcattta tggacttcta aaaaacccat ttttatctgt
     4621 ccgcttccgg acaacaacag ggcttgattc tgcagtagat cctgtccaga tgaaaaatgt
     4681 cacctttgaa catgttaaag gggtggagga agctaaacaa gaattacagg aagttgttga
     4741 attcttgaaa aatccacaaa aatttactat tcttggaggt aaacttccaa aaggaattct
     4801 tttagttgga cccccaggga ctggaaagac acttcttgcc cgagctgtgg cgggagaagc
     4861 tgatgttcct ttttattatg cttctggatc cgaatttgat gagatgtttg tgggtgtggg
     4921 agccagccgt atcagaaatc tttttaggga agcaaaggcg aatgctcctt gtgttatatt
     4981 tattgatgaa ttagattctg ttggtgggaa gagaattgaa tctccaatgc atccatattc
     5041 aaggcagacc ataaatcaac ttcttgctga aatggatggt tttaaaccca atgaaggagt
     5101 tatcataata ggagccacaa acttcccaga ggcattagat aatgccttaa tacgtcctgg
     5161 tcgttttgac atgcaagtta cagttccaag gccagatgta aaaggtcgaa cagaaatttt
     5221 gaaatggtat ctcaataaaa taaagtttga tcaatccgtt gatccagaaa ttatagctcg
     5281 aggtactgtt ggcttttccg gagcagagtt ggagaatctt gtgaaccagg ctgcattaaa
     5341 agcagctgtt gatggaaaag aaatggttac catgaaggag ctggagtttt ccaaagacaa
     5401 aattctaatg gggcctgaaa gaagaagtgt ggaaattgat aacaaaaaca aaaccatcac
     5461 agcatatcat gaatctggtc atgccattat tgcatattac acaaaagatg caatgcctat
     5521 caacaaagct acaatcatgc cacgggggcc aacacttgga catgtgtccc tgttacctga
     5581 gaatgacaga tggaatgaaa ctagagccca gctgcttgca caaatggatg ttagtatggg
     5641 aggaagagtg gcagaggagc ttatatttgg aaccgaccat attacaacag gtgcttccag
     5701 tgattttgat aatgccacta aaatagcaaa gcggatggtt accaaatttg gaatgagtga
     5761 aaagcttgga gttatgacct acagtgatac agggaaacta agtccagaaa cccaatctgc
     5821 catcgaacaa gaaataagaa tccttctaag ggactcatat gaacgagcaa aacatatctt
     5881 gaaaactcat gcaaaggagc ataagaatct cgcagaagct ttattgacct atgagacttt
     5941 ggatgccaaa gagattcaaa ttgttcttga ggggaaaaag ttggaagtga gactcgagca
     6001 ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag ctgagttggc
     6061 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     6121 gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc gccctgtagc
     6181 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     6241 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     6301 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     6361 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     6421 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     6481 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     6541 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     6601 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     6661 tttgtttatt tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa
     6721 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
     6781 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     6841 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     6901 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     6961 ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta
     7021 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
     7081 gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
     7141 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     7201 aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga
     7261 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
     7321 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     7381 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
     7441 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag
     7501 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
     7561 gacagtttta ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     7621 agaccccgta gaaaagatca aaggatcttc
//
