LOCUS       Exported                5377 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 5377)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Apr 15, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     pRSG192 designed by Brock F. Binkowski

            

            PCR amplify pF-BiBiTLc w/ BFBRSG11/BFBRSG4 (PCR #1; 2.0 kbp);

            PCR amplify pF-BiBiTsmC w/ BFBRSG5/BFBRSG9 (PCR #2; 2.8 kbp); 

            PCR amplify pF-4B w/ BFBRSG10/BFBRSG12 (464 bp);

            3X component Gibson Assembly;

            electroporation of BR610.
FEATURES             Location/Qualifiers
     source          1..5377
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    complement(15..202)
                     /label=SV40\polyA\signal
     misc_feature    complement(206..255)
                     /label=MCS 2
     misc_feature    complement(293..300)
                     /label=PmeI site
                     /note="Used in Flexi Vector Cloning"
     misc_feature    complement(301..861)
                     /label=CYBC1
                     /note="CYBC1"
     misc_feature    complement(863..870)
                     /label=SgfI Site
                     /note="Used in Flexi Vector Cloning"
     CDS             complement(871..915)
                     /codon_start=1
                     /label=15GS linker
                     /note="15GS linker"
                     /translation="GSSGGGGSGGGGSSG"
     gene            complement(916..1392)
                     /label=LgBiT
                     /note="LgBiT"
     misc_feature    1406..1432
                     /label=B globin leader
                     /note="B globin leader"
     misc_feature    complement(1433..1449)
                     /label=SP6 promoter
                     /note="SP6 promoter"
     misc_feature    complement(1452..1497)
                     /label=Minimal CMV Promoter
     misc_feature    1475..1497
                     /label=core
                     /note="core"
     promoter        1507..2061
                     /label=CMV Enhancer
                     /note="Has EcoICRI site removed. Named changed from ""CMV 
                     Promoter"" to ""CMV immediate-early enhancer/promoter"" 
                     3/21/2011 and J Hartnet's request to fit Sci Com's naming."
     promoter        2079..2145
                     /label=EM7 bacterial promoter
     CDS             2159..2953
                     /label=KanR
                     /note="Name changed frm ""Neomycin phosphotransferase"" to 
                     ""Neomycin phosphotransferase coding region"" on 3/21/2011 
                     at Jim Hartnet's request to fit Sci Com's naming."
     rep_origin      3108..3144
                     /direction=RIGHT
                     /label=ColE1-derived plasmid replication origin
                     /note="The actual origin is the first base."
     polyA_signal    complement(3826..3873)
                     /label=Synthetic PolyAdenylation Signal
     CDS             complement(3897..4934)
                     /label=HygroR
                     /note=" Feature found by back translation of amino acid 
                     feature definition."
     misc_feature    complement(4959..5377)
                     /label=SV40 promoter
                     /note="SV40 promoter"
ORIGIN
        1 aagacgcgtg acgtcggcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
       61 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
      121 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
      181 aatgtggtat ggctgattat gatccgcggc cgcaatacgt cgacgttatc agctgacttc
      241 gtacgagagc ctaggtgttt ctagaggatc cccgggtacc gagcccgaat tcgtttaaac
      301 gctctggctt gcagggtcac cggcctcact gtcgctgctc tgagacagct ctgtggggct
      361 ctcaaggcag tgcagctcca ggaagctggt gatgagcttg gcgatggctt ccacatcact
      421 gcggtggccc atgactgcac tctgcgtgag ggggtgggag aagcccgtcg caagccggag
      481 caccaccatg tagcctttcc cgaagtaccg gaccttctcc tcctccacgc tcacatcacg
      541 gacatcatgg agcaggacca ccacctggtc gtggccagct ctgaaaagag tcagcagctt
      601 cttgtagagg ctgaacgtct tcaaaacaac cttccctgtg ctcttgtcga agatggcttc
      661 ctcccagtcc tccaagttct gcacagccac aaacaggcag cctgtgacgt agaagagctt
      721 ccagcccagg ctatctccgc tgtagtaggc agcagccagg ccaatcgaca agattccaac
      781 cagcagggac caggaccgga tgcctggagc cctcttcaga tggaggcggg agctggtgcg
      841 ggtctccacc tgcaggtaca tggcgatcgc accgctcgag cctccacctc cgctcccgcc
      901 accaccggag ctcccactgt tgatggttac tcggaacagc atggagccgt cgggggtgat
      961 caggcgctcg tcgataattt tgttgccgtt ccacagggtc cctgttacag tgatcttttt
     1021 gccgtcgaac acggcgatgc cttcatacgg ccgtccgaaa tagttcagca tgttcggcgt
     1081 aaccccgtcg attaccagtg tgccataggg caggatcacc ttaaagtgat gatcatccac
     1141 agggtacacc accttaaaca cctcttcgat ctgggccatt tggtcggcgc tcagaccttc
     1201 atacgggatg atgacatgga tgtcgatctt cagggcattt tcaccgctcc ggacaatcct
     1261 ttggatcgga gttacggaca cggcgagatt ctgcagcaaa ctggacacac ctccctgttc
     1321 aaggacttgg tccaggttgt aggcggctgt ctgttcccag tccccaacga aatcttcgag
     1381 tgtgaagacc atggtgagcg ctagctgcaa aaagaacaag tagcttgtat tctatagtgt
     1441 cacctaaatg ggcgatctga cggttcacta aaccagctct gcttatatag gcctccctgg
     1501 cgcgccacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     1561 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     1621 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     1681 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     1741 catatgccaa gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     1801 gcccagtaca tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc
     1861 gctattacca tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac
     1921 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     1981 aatcaacggg actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt
     2041 aggcgtgtac ggtgggaggt caaatcgaat tctaatactg ttgacaatta atcatcggca
     2101 tagtatatcg gcatagtata atacgacaag gtgaggaact aaacccagga ggcagatcat
     2161 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     2221 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     2281 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     2341 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     2401 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     2461 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     2521 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     2581 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     2641 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     2701 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     2761 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     2821 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     2881 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     2941 cgagttcttc tgattcgaaa tgaccgacca agcgacgccc aaccggtatc agctcactca
     3001 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3061 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3121 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3181 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3241 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3301 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3361 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3421 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3481 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3541 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3601 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3661 tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag aagatccttt gatcttttct
     3721 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3781 tcaaaaagga tcttcaccta gatcctttta tagtccggaa tcgtcacaca aaaaaccaac
     3841 acacagatgt aatgaaaata aagatatttt attgcggcca tcgtgatggc tagagtctaa
     3901 actcgaccta cctccttagc gcgcggtcgt gtactgggcc ggcggttgcc gctgtcggcc
     3961 agcacctcga cgcagccgtc ggtccatacg gctgcgctcc ggcgagcgat ttgtgtgcga
     4021 ccgacggtgc cggccccgct gcggacgatg gcatcgcagc ggccttgagc ccaggcagca
     4081 tcgtcgaagt tgccgtccac gaggctctgg tagagctgat ccaggccgat gcgcagcatg
     4141 taggctcgca gacgagggct gccggccagc tcgggatgcc ggcgctcgaa gtagcgagtc
     4201 tgctgctcca tgcaagccag ccagggccgc cagaagaaga tgttggccac ctcgtactga
     4261 ctgtccccga acatagcttc ggaccagtcg attacggcgg tgatgcggcc gttgtcggtc
     4321 aggacgttgt tgctgccgaa gtcggcatgg acgaggtggc gcacctcggg acagtcttcg
     4381 gcccacagca tcagttcgtc cagggcttga gctacgctgg cggacacggt gtcgtccatc
     4441 acggtctgcc agtggtagac atggggatca gcaatggcgc aaatgaaatc ccgccaggtt
     4501 gtgtactggc cgatgccttg gggcccgaat gggccgaagc cgctggtttg actgaggtcg
     4561 gcggcggcaa tagcatccat agcttcggcg acaggctgta acacagcagg cagctctgtt
     4621 tcggggaggt cttggagagt aacgccttgg gcgcgtctac tgatgcagta tgtcaggctc
     4681 tcgctgaact cgccgatgtc caacacttcg gggatgggta gtgcagcgct ggcgaagtgg
     4741 cggtacacgt agcggtcttt gtagaagcca tcagcgcagc tattcacccg cagtacatag
     4801 ccgcgtccgc cgacatcgaa gctgaaggct cggctctctt cgccctccga caactgcatc
     4861 aggtcgctca cactgtcgaa cttctcgatg agaaattttt caacgctggt agcggtgagt
     4921 tcgggcttct tcatggtggc gctagtgtca gaagaatcga gctttttgca aaagcctagg
     4981 cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc
     5041 tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga
     5101 gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc
     5161 atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac
     5221 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
     5281 cctaactgac acacattcca cagctggttc tttccgcctc agaaggtacc taaccaagtt
     5341 cctctttcag aggttatttc aggccatggt gctgcgc
//
