LOCUS       Exported                6006 bp ds-DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6006)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Apr 15, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     pRSG428 TK N-terminal IL6-HiBiT-8GS Flexi entry vector

            

            Synthesize 789bp TK fragment using primers below and pFC32A

            Digest pRSG119 with BglII and NheI sequentially.

            Use Gibson assembly to clone and transform into BR610 cell.

            

            RSG427 tk_fwd 	cgtccggcgtagaggatcgagatctAAATGAGTCTTCGGACCTC 	 
            3'Tm=61.1 3'Ta(annealing temp)=62.1

            RSG428 tk_rev 	gccgctcaccatggtgagcgctagCGGTGGCTTTACCAACAG  	 
            3'Tm=63.6 3'Ta(annealing temp)=62.1

            

            TK_from_pAPH32_fwd 	cgtccggcgtagaggatcgaGATCTAAATGAGTCTTCGGACCTC 	 
            3'Tm=64.1 3'Ta(annealing temp)=64.6

            TK_from_pAPH32_rev 	gccgctcaccatggtgagcgctagCGGTGGCTTTACCAACAG 	 
            3'Tm=63.6 3'Ta(annealing temp)=64.6
FEATURES             Location/Qualifiers
     source          1..6006
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        6..758
                     /label=HSV-TK
                     /note="HSV-TK"
     CDS             807..842
                     /codon_start=1
                     /label=HiBiT
                     /note="HiBiT"
                     /translation="MVSGWRLFKKIS"
     CDS             843..866
                     /codon_start=1
                     /label=8GS linker
                     /note="8GS linker"
                     /translation="GSSGGSSG"
     misc_feature    867..874
                     /label=SgfI Site
                     /note="Used in Flexi Vector Cloning"
     misc_feature    876..2840
                     /label=SRPK1
                     /note="SRPK1"
     misc_feature    2841..2848
                     /label=PmeI site
                     /note="Used in Flexi Vector Cloning"
     polyA_signal    3000..3221
                     /label=Sv40 late polyadenylation signal
     promoter        3320..3738
                     /label=SV40 promoter
                     /note="Name changed frm %22SV40 early enhancer/promoter 
                     region%22 to %22SV40 enhancer and early promoter%22 on 
                     3/21/2011 at Jim Hartnet's request to fit Sci Com's 
                     naming."
     promoter        3746..3812
                     /label=EM7 bacterial promoter
     CDS             3826..4620
                     /label=NeoR / KanR
                     /note="Name changed frm %22Neomycin phosphotransferase%22 
                     to %22Neomycin phosphotransferase coding region%22 on 
                     3/21/2011 at Jim Hartnet's request to fit Sci Com's 
                     naming."
     polyA_signal    4684..4732
                     /label=Synthetic PolyAdenylation Signal
     rep_origin      4968..5004
                     /direction=RIGHT
                     /label=ColE1-derived plasmid replication origin
                     /note="The actual origin is the first base."
ORIGIN
        1 gatctaaatg agtcttcgga cctcgcgggg gccgcttaag cggtggttag ggtttgtctg
       61 acgcgggggg agggggaagg aacgaaacac tctcattcgg aggcggctcg gggtttggtc
      121 ttggtggcca cgggcacgca gaagagcgcc gcgatcctct taagcacccc cccgccctcc
      181 gtggaggcgg gggtttggtc ggcgggtggt aactggcggg ccgctgactc gggcgggtcg
      241 cgcgccccag agtgtgacct tttcggtctg ctcgcagacc cccgggcggc gccgccgcgg
      301 cggcgacggg ctcgctgggt cctaggctcc atggggaccg tatacgtgga caggctctgg
      361 agcatccgca cgactgcggt gatattaccg gagaccttct gcgggacgag ccgggtcacg
      421 cggctgacgc ggagcgtccg ttgggcgaca aacaccagga cggggcacag gtacactatc
      481 ttgtcacccg gaggcgcgag ggactgcagg agcttcaggg agtggcgcag ctgcttcatc
      541 cccgtggccc gttgctcgcg tttgctggcg gtgtccccgg aagaaatata tttgcatgtc
      601 tttagttcta tgatgacaca aaccccgccc agcgtcttgt cattggcgaa gtcgaacacg
      661 cagatgcagt cggggcggcg cggtcccagg tccacttcgc atattaaggt gacgcgtgtg
      721 gcctcgaaca ccgagcgacc ctgcagcgac ccgcttaaaa gcttggcaat ccggtactgt
      781 tggtaaagcc accgctagcg ctcaccatgg tgagcggctg gcggctgttc aagaagatta
      841 gcgggagctc cggtggctcg agcggtgcga tcgccatgga gcggaaagtg cttgcgctcc
      901 aggcccgaaa gaaaaggacc aaggccaaga aggacaaagc ccaaaggaaa tctgaaactc
      961 agcaccgagg ctctgctccc cactctgaga gtgatctacc agagcaggaa gaggagattc
     1021 tgggatctga tgatgatgag caagaagatc ctaatgatta ttgtaaagga ggttatcatc
     1081 ttgtgaaaat tggagatcta ttcaatggga gataccatgt gatccgaaag ttaggctggg
     1141 gacacttttc aacagtatgg ttatcatggg atattcaggg gaagaaattt gtggcaatga
     1201 aagtagttaa aagtgctgaa cattacactg aaacagcact agatgaaatc cggttgctga
     1261 agtcagttcg caattcagac cctaatgatc caaatagaga aatggttgtt caactactag
     1321 atgactttaa aatatcagga gttaatggaa cacatatctg catggtattt gaagttttgg
     1381 ggcatcatct gctcaagtgg atcatcaaat ccaattatca ggggcttcca ctgccttgtg
     1441 tcaaaaaaat tattcagcaa gtgttacagg gtcttgatta tttacatacc aagtgccgta
     1501 tcatccacac tgacattaaa ccagagaaca tcttattgtc agtgaatgag cagtacattc
     1561 ggaggctggc tgcagaagca acagaatggc agcgatctgg agctcctccg ccttccggat
     1621 ctgcagtcag tactgctccc cagcctaaac cagctgacaa aatgtcaaag aataagaaga
     1681 agaaattgaa gaagaagcag aagcgccagg cagaattact agagaagcga atgcaggaaa
     1741 ttgaggaaat ggagaaagag tcgggccctg ggcaaaaaag accaaacaag caagaagaat
     1801 cagagagtcc tgttgaaaga cccttgaaag agaacccacc taataaaatg acccaagaaa
     1861 aacttgaaga gtcaagtacc attggccagg atcaaacgct tatggaacgt gatacagagg
     1921 gtggtgcagc agaaattaat tgcaatggag tgattgaagt cattaattat actcagaaca
     1981 gtaataatga aacattgaga cataaagagg atctacataa tgctaatgac tgtgatgtcc
     2041 aaaatttgaa tcaggaatct agtttcctaa gctcccaaaa tggagacagc agcacatctc
     2101 aagaaacaga ctcttgtaca cctataacat ctgaggtgtc agacaccatg gtgtgccagt
     2161 cttcctcaac tgtaggtcag tcattcagtg aacaacacat tagccaactt caagaaagca
     2221 ttcgggcaga gataccctgt gaagatgaac aagagcaaga acataacgga ccactggaca
     2281 acaaaggaaa atccacggct ggaaattttc ttgttaatcc ccttgagcca aaaaatgcag
     2341 aaaagctcaa ggtgaagatt gctgaccttg gaaatgcttg ttgggtgcac aaacatttca
     2401 ctgaagatat tcaaacaagg caatatcgtt ccttggaagt tctaatcgga tctggctata
     2461 atacccctgc tgacatttgg agcacggcat gcatggcctt tgaactggcc acaggtgact
     2521 atttgtttga acctcattca ggggaagagt acactcgaga tgaagatcac attgcattga
     2581 tcatagaact tctggggaag gtgcctcgca agctcattgt ggcaggaaaa tattccaagg
     2641 aatttttcac caaaaaaggt gacctgaaac atatcacgaa gctgaaacct tggggccttt
     2701 ttgaggttct agtggagaag tatgagtggt cgcaggaaga ggcagctggc ttcacagatt
     2761 tcttactgcc catgttggag ctgatccctg agaagagagc cactgccgcc gagtgtctcc
     2821 ggcacccttg gcttaactcc gtttaaacga attcgggctc ggtacccggg gatcctctag
     2881 agtcgacctg caggcatgca agctgatccg gctgctaaca aagcccgaaa ggaagctgag
     2941 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcggc cgcttcgagc
     3001 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     3061 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     3121 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg
     3181 ggaggttttt ttaagcaagt aaaacctcta caaatgtggt aaaatcgaat tttaacaaaa
     3241 tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     3301 acaccgcata cgcggatctg cgcagcacca tggcctgaaa taacctctga aagaggaact
     3361 tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     3421 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3481 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3541 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3601 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3661 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3721 ggcttttgca aaaagcttaa ttaactgttg acaattaatc atcggcatag tatatcggca
     3781 tagtataata cgacaaggtg aggaactaaa cccaggaggc agatcatgat tgaacaagat
     3841 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3901 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3961 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4021 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4081 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4141 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4201 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4261 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4321 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4381 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4441 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4501 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4561 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4621 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgatggc
     4681 cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata
     4741 gcgataagga tcctctttgc gcttgcgttt tcccttgtcc agatagccca gtagctgaca
     4801 ttcatccggg gtcagcaccg tttctgcgga ctggctttct acccggtatc agctcactca
     4861 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4921 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4981 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5041 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5101 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5161 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5221 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5281 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5341 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5401 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5461 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5521 tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag aagatccttt gatcttttct
     5581 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5641 tcaaaaagga tcttcaccta gatcctttta tagtccggaa atacaggaac gcacgctgga
     5701 tggcccttcg ctgggatggt gaaaccatga aaaatggcag cttcagtgga ttaagtgggg
     5761 gtaatgtggc ctgtaccctc tggttgcata ggtattcata cggttaaaat ttatcaggcg
     5821 cgattgcggc agtttttcgg gtggtttgtt gccattttta cctgtctgct gccgtgatcg
     5881 cgctgaacgc gttttagcgg tgcgtacaat taagggatta tggtaaatcc acttactgtc
     5941 tgccctcgta gccatcgaga taaaccgcag tactccggcc acgatgcgtc cggcgtagag
     6001 gatcga
//
