LOCUS       T7-LacO-HaloTag-        5303 bp DNA     circular SYN 15-FEB-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5303)
  AUTHORS   Stanford University
  TITLE     Direct Submission
  JOURNAL   Exported Feb 15, 2024 from SnapGene 6.2.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5303
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      578..1166
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1436..1452
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1915..1933
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1934..1958
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator 
                     toinhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     misc_feature    2001..2904
                     /label=HaloTag
     CDS             2910..2960
                     /codon_start=1
                     /label=RGD
                     /translation="TVYAVTGRGDSPASSAA"
     CDS             2970..3202
                     /codon_start=1
                     /label=TitinInsert2910-1
                     /translation="METLHITKTMKNIEVPETKTASFECEVSHFNVPSMWLKNGVEIEM
                     SEKFKIVVQGKLHQLIIMNTSTEDSAEYTFVC"
     CDS             3204..3236
                     /codon_start=1
                     /label=HiBiT
                     /translation="VSGWRLFKKIS"
     CDS             3237..3268
                     /codon_start=1
                     /label=TitinInsert2910-1
                     /translation="NDQVSATLTV"
     CDS             3297..3314
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      3322..3369
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3678..3769
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3770..4630
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             join(4713..5303,1..492)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator 
                     toinhibit 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"
ORIGIN
        1 ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa
       61 ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc
      121 gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt
      181 accgagtccg ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa
      241 gatagctcat gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg
      301 caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag
      361 ctgttgccag tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc
      421 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
      481 agcgggcagt gactcatgac caaaatccct taacgtgagt tacgcgcgcg tcgttccact
      541 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
      601 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
      661 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
      721 ctgttcttct agtgtagccg tagttagccc accacttcaa gaactctgta gcaccgccta
      781 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
      841 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
      901 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
      961 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     1021 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     1081 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     1141 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     1201 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     1261 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     1321 gcgagtcagt gagcgaggaa gcggaaggcg agagtaggga actgccaggc atcaaactaa
     1381 gcagaaggcc cctgacggat ggcctttttg cgtttctaca aactctttct gtgttgtaaa
     1441 acgacggcca gtcttaagct cgggccccct gggcggttct gataacgagt aatcgttaat
     1501 ccgcaaataa cgtaaaaacc cgcttcggcg ggttttttta tggggggagt ttagggaaag
     1561 agcatttgtc agaatattta agggcgcctg tcactttgct tgatatatga gaattattta
     1621 accttataaa tgagaaaaaa gcaacgcact ttaaataaga tacgttgctt tttcgattga
     1681 tgaacaccta taattaaact attcatctat tatttatgat tttttgtata tacaatattt
     1741 ctagtttgtt aaagagaatt aagaaaataa atctcgaaaa taataaaggg aaaatcagtt
     1801 tttgatatca aaattataca tgtcaacgat aatacaaaat ataatacaaa ctataagatg
     1861 ttatcagtat ttattatcat ttagaataaa ttttgtgtcg cccttccgcg aaattaatac
     1921 gactcactat aggggaattg tgagcggata acaattcccc tctagaaata attttgttta
     1981 acttttagga ggtaaaacat atgggcagcg aaattggcac tggttttccg tttgatccgc
     2041 attacgttga agttttaggc gaacgcatgc actacgtaga cgtgggtccg cgtgacggca
     2101 ccccggtcct gttcctgcac ggcaatccaa cctctagcta tgtctggcgc aacattatcc
     2161 cgcacgtcgc gccgacgcac cgtagtattg caccggatct gatcggtatg ggtaaaagcg
     2221 ataaaccgga cctgggctat ttctttgacg accacgttcg ttttatggat gcgttcattg
     2281 aggccttggg tctggaagag gtggtgttgg ttatccatga ctggggtagc gctctgggct
     2341 tccactgggc gaagcgcaac ccggaacgtg tgaagggcat cgcatttatg gagttcattc
     2401 gcccgattcc gacctgggac gagtggccgg agttcgcgcg tgaaacgttc caggcctttc
     2461 gcaccaccga tgtcggtcgt aagctgatta ttgaccaaaa tgttttcatc gagggcacgt
     2521 tgccgatggg cgttgttcgt ccgctgaccg aagtcgagat ggaccattac cgtgagccgt
     2581 ttttgaatcc ggtggatcgt gagccgctgt ggcgtttccc gaacgagctg ccgatcgccg
     2641 gtgaaccggc gaacatcgtt gcgctggttg aagagtacat ggactggctg caccagagcc
     2701 cggtgccgaa actgctgttt tggggtacgc cgggtgtgct gatcccgcct gcggaagcag
     2761 cccgcctggc gaaatccctg ccgaacgcga aggcagtgga tattggtcca ggcctgaatc
     2821 tgctgcagga agataacccg gacctgattg gtagcgaaat cgcgcgctgg ctgagcacgt
     2881 tggagatcag cggcggtgct ggcgagttca ccgtctatgc cgtcaccggt cgtggcgatt
     2941 ccccggcaag ctcggcggca ggtggcggta tggaaactct gcatattacc aaaaccatga
     3001 aaaacattga agtgccggaa accaaaaccg cgagctttga atgcgaagtg agccatttta
     3061 acgtgccgag catgtggctg aaaaacggcg tggaaattga aatgagcgaa aaatttaaaa
     3121 ttgtggtgca gggcaaactg catcagctga ttattatgaa caccagcacc gaagatagcg
     3181 cggaatatac ctttgtgtgc ggtgttagtg gctggcgtct gtttaagaaa attagcaacg
     3241 atcaggtgag cgcgaccctg accgtgactt ccgagaatct gtacttcgag caaggtcacc
     3301 accatcatca ccattaaccc cctagcataa ccccttgggg cctctaaacg ggtcttgagg
     3361 ggttttttgc ccctgagacg cgtcaatcga gttcgtacct aagggcgaca ccccctaatt
     3421 agcccgggcg aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctggcag
     3481 ttccctactc tcgcatgggg agtccccaca ctaccatcgg cgctacggcg tttcacttct
     3541 gagttcggca tggggtcagg tgggaccacc gcgctactgc cgccaggcaa acaaggggtg
     3601 ttatgagcca tattcaggta taaatgggct cgcgataatg ttcagaattg gttaattggt
     3661 tgtaacactg acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3721 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagaata tgagtattca
     3781 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3841 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     3901 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     3961 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4021 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4081 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4141 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4201 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4261 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcgat
     4321 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4381 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4441 ggctggctgg tttattgctg ataaatccgg agccggtgag cgtggttctc gcggtatcat
     4501 cgcagcgctg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4561 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4621 gcattggtaa gcggcgcgcc atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc
     4681 gcccggaaga gagtcaattc agggtggtga atatgaaacc agtaacgtta tacgatgtcg
     4741 cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg
     4801 tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca
     4861 accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca
     4921 gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac
     4981 tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg
     5041 cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg
     5101 accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg
     5161 tctctgacca gacacccatc aacagtatta ttttctccca tgaggacggt acgcgactgg
     5221 gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa
     5281 gttctgtctc ggcgcgtctg cgt
//
