LOCUS       Exported                6515 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    CBP-HAT-GST
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6515)
  AUTHORS   Shen WF, Krishnan K, Lawrence HJ, Largman C
  TITLE     The HOX homeodomain proteins block CBP histone acetyltransferase 
            activity
  JOURNAL   Mol Cell Biol. 2001 Nov;21(21):7509-22
  PUBMED    11585930
REFERENCE   2  (bases 1 to 6515)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6515
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     56..73
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        146..223
                     /gene="lacI"
                     /label=lacI promoter
     CDS             224..1306
                     /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"
     primer_bind     complement(243..262)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        1355..1385
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1393..1409
                     /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)."
     primer_bind     1417..1433
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1417..1433
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1429..1956
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1449..1466)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1449..1465)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1458..1480)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        2058..2086
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2094..2110
                     /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)."
     CDS             2133..2786
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2744..2766
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2793..2810
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     regulatory      4092..4101
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     complement(4440..4462)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4607..4625)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4693..4797
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4798..5658
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5016..5035)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5829..6417
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6318..6337
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
       61 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
      121 cggtatttca caccgcataa attccgacac catcgaatgg cgcaaaacct ttcgcggtat
      181 ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt
      241 atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca
      301 ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa
      361 ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt
      421 tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg
      481 cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc
      541 ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta
      601 tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt
      661 atttcttgat gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg
      721 tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc
      781 gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac
      841 tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt
      901 tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa
      961 cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc
     1021 ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt
     1081 aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca
     1141 actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag
     1201 aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     1261 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     1321 atgtaagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     1381 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     1441 acggattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     1501 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     1561 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgctt tgcctggttt
     1621 ccggcaccag aagcggtgcc ggaaagctgg ctggagtgcg atcttcctga ggccgatact
     1681 gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg cgcccatcta caccaacgtg
     1741 acctatccca ttacggtcaa tccgccgttt gttcccacgg agaatccgac gggttgttac
     1801 tcgctcacat ttaatgttga tgaaagctgg ctacaggaag gccagacgcg aattattttt
     1861 gatggcgttg gaattagctt atcgactgca cggtgcacca atgcttctgg cgtcaggcag
     1921 ccatcggaag ctgtggtatg gctgtgcagg tcgtaaatca ctgcataatt cgtgtcgctc
     1981 aaggcgcact cccgttctgg ataatgtttt ttgcgccgac atcataacgg ttctggcaaa
     2041 tattctgaaa tgagctgttg acaattaatc atcggctcgt ataatgtgtg gaattgtgag
     2101 cggataacaa tttcacacag gaaacagtat tcatgtcccc tatactaggt tattggaaaa
     2161 ttaagggcct tgtgcaaccc actcgacttc ttttggaata tcttgaagaa aaatatgaag
     2221 agcatttgta tgagcgcgat gaaggtgata aatggcgaaa caaaaagttt gaattgggtt
     2281 tggagtttcc caatcttcct tattatattg atggtgatgt taaattaaca cagtctatgg
     2341 ccatcatacg ttatatagct gacaagcaca acatgttggg tggttgtcca aaagagcgtg
     2401 cagagatttc aatgcttgaa ggagcggttt tggatattag atacggtgtt tcgagaattg
     2461 catatagtaa agactttgaa actctcaaag ttgattttct tagcaagcta cctgaaatgc
     2521 tgaaaatgtt cgaagatcgt ttatgtcata aaacatattt aaatggtgat catgtaaccc
     2581 atcctgactt catgttgtat gacgctcttg atgttgtttt atacatggac ccaatgtgcc
     2641 tggatgcgtt cccaaaatta gtttgtttta aaaaacgtat tgaagctatc ccacaaattg
     2701 ataagtactt gaaatccagc aagtatatag catggccttt gcagggctgg caagccacgt
     2761 ttggtggtgg cgaccatcct ccaaaatcgg atctggttcc gcgtggatcc cttggatatt
     2821 gctgtggacg aaagtatgag ttctccccac agactttgtg ctgttacgga aagcagctgt
     2881 gtacaattcc tcgtgatgca gcctactaca gctatcagaa taggtatcat ttctgtggga
     2941 agtgtttcac agagatccag ggcgagaatg tgaccctggg tgacgaccct tcccaacctc
     3001 agacgacaat ttccaaggat caatttgaaa agaagaaaaa tgatacctta gatcctgaac
     3061 cttttgttga ctgcaaagag tgtggccgga agatgcatca gatttgtgtt ctacactatg
     3121 acatcatttg gccttcaggt tttgtgtgtg acaactgttt gaagaaaact ggcagacctc
     3181 ggaaagaaaa caaattcagt gctaagaggc tgcagaccac acgattggga aaccacttag
     3241 aagacagagt gaataagttt ttgcggcgcc agaatcaccc tgaagctggg gaggtttttg
     3301 tcagagtggt ggccagctca gacaagactg tggaggtcaa gccgggaatg aagtcaaggt
     3361 ttgtggattc tggagagatg tcggaatctt tcccatatcg taccaaagca ctctttgctt
     3421 ttgaggagat cgatggagtc gatgtgtgct tttttgggat gcatgtgcaa gaatacggct
     3481 ctgattgccc cccaccaaat acaaggcgtg tatacatatc ttatctggac agtattcatt
     3541 tcttccggcc ccgctgcctc cggacagctg tttaccatga gatcctcatc ggatatctcg
     3601 agtatgtgaa gaaattgggg tatgtgacag gacatatttg ggcctgtccc ccaagtgaag
     3661 gagatgacta tatctttcat tgccaccccc ctgaccagaa aatccccaaa ccaaaacgac
     3721 tacaggagtg gtacaagaag atgctggaca aggcgtttgc agagaggatc attaacgact
     3781 ataaggacat cttcaaacaa gcgaacgaag acaggctcac gagtgccaag gagttgccct
     3841 attttgaagg agatttctgg cctaatgtgt tggaagaaag cattaaggaa ctagaacaag
     3901 aagaagaaga aaggaaaaaa gaagagagta ctgcagcgag tgagactcct gagggcagtc
     3961 agggtgacag caaaaatgcg aagaaaaaga acaacaagaa gaccaacaaa aacaaaagca
     4021 gcattagccg cgccaacaag aagaagccca gcatgcccaa tgtttccaac gacctgtcgc
     4081 agaagctgta tgccaccatg gagaagcaca aggaggtatt ctttgtgatt catctgcatg
     4141 ctgggcctgt tatcagcact cagcccccca tcgtggaccc tgatcctctg cttagctgtg
     4201 acctcatgga tgggcgagat gccttcctca ccctggccag agacaagcac tgggaattct
     4261 cttccttacg ccgctccaaa tggtccactc tgtgcatgct ggtggagctg cacacacagg
     4321 gccaggaccg ctttgtttat acctgcaatg agtgcaaaca ccatgtggaa acacgctgag
     4381 cggccgcatc gtgactgact gacgatctgc ctcgcgcgtt tcggtgatga cggtgaaaac
     4441 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     4501 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc
     4561 cagtcacgta gcgatagcgg agtgtataat tcttgaagac gaaagggcct cgtgatacgc
     4621 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     4681 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4741 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4801 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4861 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4921 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4981 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5041 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5101 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5161 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5221 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5281 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5341 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5401 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5461 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5521 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5581 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5641 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5701 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5761 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5821 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5881 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5941 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     6001 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6061 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6121 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6181 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6241 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6301 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6361 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6421 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     6481 tttcctgcgt tatcccctga ttctgtggat aaccg
//
