LOCUS       Exported                6535 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pGEX4T-1_WT_OCT4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6535)
  AUTHORS   Brumbaugh J, Hou Z, Russell JD, Howden SE, Yu P, Ledvina AR, Coon 
            JJ, Thomson JA
  TITLE     Phosphorylation regulates human OCT4.
  JOURNAL   Proc Natl Acad Sci U S A. 2012 May 8;109(19):7162-8. Epub 2012 Apr 
            2.
  PUBMED    22474382
REFERENCE   2  (bases 1 to 6535)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6535
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..78
                     /gene="lacI"
                     /label=lacI promoter
     CDS             79..1161
                     /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(98..117)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        1210..1240
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1248..1264
                     /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     1272..1288
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1272..1288
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1284..1811
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1304..1321)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1304..1320)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1313..1335)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        1913..1941
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1949..1965
                     /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             1988..2641
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2599..2621
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2648..2665
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2672..2692
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3822..3842
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3849..3866
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3919..3937)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3933..3980
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4104..4123)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4315..4337)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4482..4500)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4568..4672
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4673..5533
                     /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(4891..4910)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5704..6292
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6193..6212
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6446..6463
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gacaccatcg aatggcgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt
       61 caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt
      121 gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg
      181 cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa
      241 caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac
      301 gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg
      361 gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt
      421 ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt
      481 gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca
      541 cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg
      601 gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg
      661 cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg
      721 gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat
      781 gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg
      841 cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac
      901 gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc
      961 ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag
     1021 ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg
     1081 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     1141 cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagctca ctcattaggc
     1201 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     1261 acaatttcac acaggaaaca gctatgacca tgattacgga ttcactggcc gtcgttttac
     1321 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     1381 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     1441 gcagcctgaa tggcgaatgg cgctttgcct ggtttccggc accagaagcg gtgccggaaa
     1501 gctggctgga gtgcgatctt cctgaggccg atactgtcgt cgtcccctca aactggcaga
     1561 tgcacggtta cgatgcgccc atctacacca acgtgaccta tcccattacg gtcaatccgc
     1621 cgtttgttcc cacggagaat ccgacgggtt gttactcgct cacatttaat gttgatgaaa
     1681 gctggctaca ggaaggccag acgcgaatta tttttgatgg cgttggaatt agcttatcga
     1741 ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg gtatggctgt
     1801 gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt tctggataat
     1861 gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc tgttgacaat
     1921 taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     1981 agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc aacccactcg
     2041 acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc gcgatgaagg
     2101 tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc ttccttatta
     2161 tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata tagctgacaa
     2221 gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc ttgaaggagc
     2281 ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact ttgaaactct
     2341 caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag atcgtttatg
     2401 tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt tgtatgacgc
     2461 tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa aattagtttg
     2521 ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat ccagcaagta
     2581 tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc atcctccaaa
     2641 atcggatctg gttccgcgtg gatctcatat ggaaaacttg tatttccagg gccatatggc
     2701 tagcgcggga cacctggctt cggatttcgc cttctcgccc cctccaggtg gtggaggtga
     2761 tgggccaggg gggccggagc cgggctgggt tgatcctcgg acctggctaa gcttccaagg
     2821 ccctcctgga gggccaggaa tcgggccggg ggttgggcca ggctctgagg tgtgggggat
     2881 tcccccatgc cccccgccgt atgagttctg tggggggatg gcgtactgtg ggccccaggt
     2941 tggagtgggg ctagtgcccc aaggcggctt ggagacctct cagcctgagg gtgaagcagg
     3001 agtcggggtg gagagcaact ccgatggggc ctccccggag ccctgcaccg tcacccctgg
     3061 tgccgtgaag ctggagaagg agaagctgga gcaaaacccg gaggagtccc aggacatcaa
     3121 agctctgcag aaagaactcg agcaatttgc caagctcctg aagcagaaga ggatcaccct
     3181 gggatataca caggccgatg tggggctcac cctgggggtt ctatttggga aggtattcag
     3241 ccaaacgacc atctgccgct ttgaggctct gcagcttagc ttcaagaaca tgtgtaagct
     3301 gcggcccttg ctgcagaagt gggtggagga agctgacaac aatgaaaatc ttcaggagat
     3361 atgcaaagca gaaaccctcg tgcaggcccg aaagagaaag cgaaccagta tcgagaaccg
     3421 agtgagaggc aacctggaga atttgttcct gcagtgcccg aaacccacac tgcagcagat
     3481 cagccacatc gcccagcagc ttgggctcga gaaggatgtg gtccgagtgt ggttctgtaa
     3541 ccggcgccag aagggcaagc gatcaagcag cgactatgca caacgagagg attttgaggc
     3601 tgctgggtct cctttctcag ggggaccagt gtcctttcct ctggccccag ggccccattt
     3661 tggtacccca ggctatggga gccctcactt cactgcactg tactcctcgg tccctttccc
     3721 tgagggggaa gcctttcccc ctgtctccgt caccactctg ggctctccca tgcattcaaa
     3781 ctgagaattc gagctccgtc gacaagctta cggccgcact cgagaacctg tacttccagg
     3841 gagtcgagca ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag
     3901 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
     3961 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc
     4021 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     4081 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     4141 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     4201 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacggg ggatcccaat
     4261 tcatcgtgac tgactgacga tctgcctcgc gcgtttcggt gatgacggtg aaaacctctg
     4321 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     4381 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc
     4441 acgtagcgat agcggagtgt ataattcttg aagacgaaag ggcctcgtga tacgcctatt
     4501 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     4561 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4621 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     4681 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4741 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     4801 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4861 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
     4921 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4981 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5041 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     5101 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5161 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc
     5221 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     5281 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5341 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5401 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5461 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5521 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     5581 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5641 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5701 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5761 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5821 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     5881 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5941 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6001 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6061 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6121 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6181 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6241 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6301 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     6361 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     6421 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     6481 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcataa attcc
//
