LOCUS       sequence-177114-        7543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7543)
  AUTHORS   Resnick RJ, Taylor SJ, Lin Q, Shalloway D
  TITLE     Phosphorylation of the Src substrate Sam68 by Cdc2 during mitosis.
  JOURNAL   Oncogene. 1997 Sep;15(11):1247-53.
  PUBMED    9315091
REFERENCE   2  (bases 1 to 7543)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Oncogene.";
            date: "1997-09"; volume: "15(11)"; pages: "1247-53"
COMMENT     Sequence Label: pGEX-2T Sam68
FEATURES             Location/Qualifiers
     source          1..7543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(51..56,57..72,73..79)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    87..103
                     /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             126..779
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     737..759
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             786..803
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(3461..3483)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3628..3646)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3714..3818
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3819..3887,3888..4679)
                     /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(4037..4056)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4850..5438
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5339..5358
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5592..5609
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5682..5759
                     /gene="lacI"
                     /label=lacI promoter
     CDS             5760..6842
                     /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(5779..5798)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6891..6896,6897..6914,6915..6921)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6929..6945
                     /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     6953..6969
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6953..6969
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             6965..7492
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(6985..7002)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6985..7001)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6994..7016)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 tttttgcgcc gacatcataa cggttctggc aaatattctg aaatgagctg ttgacaatta
       61 atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca caggaaacag
      121 tattcatgtc ccctatacta ggttattgga aaattaaggg ccttgtgcaa cccactcgac
      181 ttcttttgga atatcttgaa gaaaaatatg aagagcattt gtatgagcgc gatgaaggtg
      241 ataaatggcg aaacaaaaag tttgaattgg gtttggagtt tcccaatctt ccttattata
      301 ttgatggtga tgttaaatta acacagtcta tggccatcat acgttatata gctgacaagc
      361 acaacatgtt gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt gaaggagcgg
      421 ttttggatat tagatacggt gtttcgagaa ttgcatatag taaagacttt gaaactctca
      481 aagttgattt tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat cgtttatgtc
      541 ataaaacata tttaaatggt gatcatgtaa cccatcctga cttcatgttg tatgacgctc
      601 ttgatgttgt tttatacatg gacccaatgt gcctggatgc gttcccaaaa ttagtttgtt
      661 ttaaaaaacg tattgaagct atcccacaaa ttgataagta cttgaaatcc agcaagtata
      721 tagcatggcc tttgcagggc tggcaagcca cgtttggtgg tggcgaccat cctccaaaat
      781 cggatctggt tccgcgtgga tcccagcgcc gggacgatcc tgcctcgcgc ctcacccggt
      841 cctcgggccg cagctgctcc aaggacccgt caggtgccca cccctcggtg cgtctgaccc
      901 cgtctcggcc gtcgccgctt cctcaccggc cccggggagg gggaggtggg cccagaggag
      961 gcgctcgggc ctcgcccgcc acccagccgc cgccgctgct gcctccctcc acccctggtc
     1021 ccgacgcgac ggtggtgggt tccgcgccga ccccgctgct gcccccgtca gccacagccg
     1081 cggtcaagat ggagccggag aataagtacc tgcctgaact catggccgag aaggactcgc
     1141 tcgacccgtc cttcactcac gccatgcagc tgctgtccgt agaaattgag aagattcaga
     1201 agggagagtc aaaaaaagat gacgaggaga attatttgga tttattttct cataagaaca
     1261 tgaagctgaa agaacgcgtg ctgatacctg tcaagcagta tccaaagttc aattttgtgg
     1321 ggaagattct tggaccacaa ggaaatacaa tcaaaagact ccaggaagag actggtgcaa
     1381 agatctctgt cttggggaag ggttcaatga gagacaaagc caaggaggaa gagttgcgca
     1441 agggtggaga ccccaaatat gcccatttaa atatggatct gcatgtcttc attgaagtct
     1501 ttggaccccc gtgtgaagct tatgctctta tggcccatgc tatggaagaa gtcaagaagt
     1561 tcctagtacc agatatgatg gatgatatct gtcaggagca gtttctagaa ttgtcctact
     1621 tgaacggagt acctgaaccc tctcgtggtc gtggggtatc tgtgagagga cgaggagctg
     1681 cccctcctcc tccacctgtt cccagaggac gtggtgttgg accacctaga ggagctttgg
     1741 ttcgtggaac cccagtgaga ggctccatca ccagaggtgc cactgtgact cgaggagtgc
     1801 cacccccacc tactgtgagg ggtgctccaa caccaagagc tcggacagct gggattcaga
     1861 gaataccttt gcctcccaca cctgcaccag aaacatacga agattatgga tatgatgata
     1921 catacgcaga acagagttac gaaggctatg aaggctatta cagccagagt caaggggagt
     1981 cagagtatta tgactatgga catggggagc tccaagattc ttacgaagcc tacggacaag
     2041 atgactggaa tgggaccagg ccatcactga aggctcctcc agctaggcca gtgaagggag
     2101 catacagaga gcatccatat ggacgttatt aaaaacaaac aggagggaaa aatatcagtt
     2161 atgagcaaag ttgttactga tttcttgtat cccaggattc ctgttgcttt acccacaaca
     2221 gacaagtaat tgtctaagtg tttttcttcg tggtcccttt cttttcccca cttcctccat
     2281 tcttaactct gcattctggc ttctgtatgt agtattttaa aatgagttaa aatagattta
     2341 ggaatatcga attaattttt taagtgtgta gatgcttttt tttctttgtt gtttaaatat
     2401 aaacagtgta ccttttataa taaaaaaaag ttgagttaaa aaaaaaccac aaacatgtta
     2461 gtttcaaaag tggcattgct tgcttaaagg ttctgaaggc ttgttaactt tttcttagtt
     2521 ttgatttgag gcaccccgta aagtcatagt tgcagaatcc caaactaggc tacatttcaa
     2581 aattcagggc tgcttaagat gtaaaaccac aatgttaaca acggcagtag tgccaccatg
     2641 taaaagtgag ctcagaggtc tctaaaacat ttcctttaaa agcacatggc tgttgaatct
     2701 taaggttaaa ttttaatatg aaagctcctc atgaattaaa tagttgatgc aatttttaac
     2761 gttaattgac ttaaaacaac aacaaaatta ggtttgtaaa agtgactttt tcattatgtg
     2821 ggttttgaaa tctagcccca gacatacagt gttgagagat acttagtggg ggaggggtgg
     2881 gctctgaata agattgatgg ggagagaagc tagccaccta aatggaatct gatggagttc
     2941 tttttagcca caatcttttc aataatagta ctaaaaatca aattttcagt atttaaaatg
     3001 tcaagagtat tttgtccatc tgagattcta cactccatga aaaactcact tggacactgg
     3061 ggccaaaagc tgatgatgtt caataaattg accattgtca gttttgagct gttcccaagg
     3121 tagaaggcag ttaagtatgt ctcaacacta gcatgatata aaaagggaca ctgcagctga
     3181 atgagaaaag aatcaaaatc cactttgtac ataagttaaa gtcctaatgg atttgtaccg
     3241 tcctcccatt ttgttttcgg aagattaaat gctacatgtg taagtctgcc taaataggta
     3301 gcttaaactt atgtcaaaat gtctgcagca gtttgtcaat aaagtttagc tcgtgccgaa
     3361 ttcctgcagc ccgggagggg gaggtgggcc cagaggaaag ccgaattcat cgtgactgac
     3421 tgacgatctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     3481 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     3541 cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg
     3601 gagtgtataa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     3661 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3721 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3781 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     3841 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     3901 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     3961 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     4021 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc
     4081 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     4141 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4201 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4261 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     4321 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt
     4381 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     4441 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     4501 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     4561 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     4621 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     4681 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4741 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4801 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4861 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4921 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4981 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5041 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5101 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5161 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5221 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5281 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5341 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5401 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     5461 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     5521 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     5581 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
     5641 tccttacgca tctgtgcggt atttcacacc gcataaattc cgacaccatc gaatggcgca
     5701 aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg gtggtgaatg
     5761 tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat cagaccgttt
     5821 cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg
     5881 cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg ggcaaacagt
     5941 cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg caaattgtcg
     6001 cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac
     6061 gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg
     6121 ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa gctgcctgca
     6181 ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac agtattattt
     6241 tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg ggtcaccagc
     6301 aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct
     6361 ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga
     6421 gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg
     6481 cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg
     6541 ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa gacagctcat
     6601 gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg
     6661 tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg
     6721 tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg
     6781 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     6841 gagcgcaacg caattaatgt aagttagctc actcattagg caccccaggc tttacacttt
     6901 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     6961 agctatgacc atgattacgg attcactggc cgtcgtttta caacgtcgtg actgggaaaa
     7021 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     7081 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     7141 gcgctttgcc tggtttccgg caccagaagc ggtgccggaa agctggctgg agtgcgatct
     7201 tcctgaggcc gatactgtcg tcgtcccctc aaactggcag atgcacggtt acgatgcgcc
     7261 catctacacc aacgtgacct atcccattac ggtcaatccg ccgtttgttc ccacggagaa
     7321 tccgacgggt tgttactcgc tcacatttaa tgttgatgaa agctggctac aggaaggcca
     7381 gacgcgaatt atttttgatg gcgttggaat tagcttatcg actgcacggt gcaccaatgc
     7441 ttctggcgtc aggcagccat cggaagctgt ggtatggctg tgcaggtcgt aaatcactgc
     7501 ataattcgtg tcgctcaagg cgcactcccg ttctggataa tgt
//
