LOCUS       Exported                5480 bp DNA     circular SYN 23-JUL-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5480)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 8, 2023 from SnapGene Viewer 6.0.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5480
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     577..603
                     /label=3578F
     primer_bind     578..604
                     /label=3580f
     regulatory      597..606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     599..617
                     /label=3522F-HindIII-BTB
     CDS             603..989
                     /codon_start=1
                     /label=BTB
                     /translation="MASPADSCIQFTRHASDVLLNLNRLRSRDILTDVVIVVSREQFRA
                     HKTVLMACSGLFYSIFTDQLKCNLSVINLDPEINPEGFCILLDFMYTSRLNLREGNIMA
                     VMATAMYLQMEHVVDTCRKFIKASE"
     primer_bind     complement(966..991)
                     /label=3523R-BTB-BamHI
     primer_bind     1001..1043
                     /label=3522F-HindIII-BTB
     CDS             1029..1415
                     /codon_start=1
                     /label=BTB
                     /translation="MASPADSCIQFTRHASDVLLNLNRLRSRDILTDVVIVVSREQFRA
                     HKTVLMACSGLFYSIFTDQLKCNLSVINLDPEINPEGFCILLDFMYTSRLNLREGNIMA
                     VMATAMYLQMEHVVDTCRKFIKASE"
     primer_bind     complement(1392..1427)
                     /label=3523R-BTB-BamHI
     primer_bind     complement(1413..1433)
                     /label=3579R
     primer_bind     complement(1414..1433)
                     /label=3581
     regulatory      1431..1440
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1437..2147
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     polyA_signal    2268..2389
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2396..2851)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2878..2982
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2984..3341
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3192..3327
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3376..4170
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4402..4449
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4778..5366
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcca
      601 ccatggcctc gccggctgac agctgtatcc agttcacccg ccatgccagt gatgttcttc
      661 tcaaccttaa tcgtctccgg agtcgagaca tcttgactga tgttgtcatt gttgtgagcc
      721 gtgagcagtt tagagcccat aaaacggtcc tcatggcctg cagtggcctg ttctatagca
      781 tctttacaga ccagttgaaa tgcaacctta gtgtgatcaa tctagatcct gagatcaacc
      841 ctgagggatt ctgcatcctc ctggacttca tgtacacatc tcggctcaat ttgcgggagg
      901 gcaacatcat ggctgtgatg gccacggcta tgtacctgca gatggagcat gttgtggaca
      961 cttgccggaa gtttattaag gccagtgaag gcggaagcgg cggaaagctt ggcggaagcg
     1021 gaggcagcat ggcctcgccg gctgacagct gtatccagtt cacccgccat gccagtgatg
     1081 ttcttctcaa ccttaatcgt ctccggagtc gagacatctt gactgatgtt gtcattgttg
     1141 tgagccgtga gcagtttaga gcccataaaa cggtcctcat ggcctgcagt ggcctgttct
     1201 atagcatctt tacagaccag ttgaaatgca accttagtgt gatcaatcta gatcctgaga
     1261 tcaaccctga gggattctgc atcctcctgg acttcatgta cacatctcgg ctcaatttgc
     1321 gggagggcaa catcatggct gtgatggcca cggctatgta cctgcagatg gagcatgttg
     1381 tggacacttg ccggaagttt attaaggcca gtgaagggga tccaccggtc gccaccatgg
     1441 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
     1501 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     1561 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
     1621 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
     1681 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     1741 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg
     1801 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
     1861 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
     1921 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     1981 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     2041 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     2101 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtagcgg ccgcgactct
     2161 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     2221 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     2281 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     2341 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
     2401 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     2461 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     2521 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     2581 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     2641 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     2701 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     2761 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     2821 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     2881 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2941 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     3001 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3061 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3121 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3181 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3241 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3301 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     3361 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3421 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3481 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3541 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3601 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3661 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3721 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3781 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     3841 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     3901 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     3961 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4021 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4081 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4141 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     4201 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4261 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4321 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     4381 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     4441 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     4501 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     4561 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     4621 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4681 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4741 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4801 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4861 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4921 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4981 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5041 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5101 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5161 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5221 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5281 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5341 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5401 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     5461 accgtattac cgccatgcat
//
