LOCUS       Exported                4870 bp ds-DNA     circular SYN 25-SEP-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4870)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 25, 2023 from SnapGene 4.1.8
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4870
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     348..367
                     /label=Seq-F
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    563..589
                     /label=primer1
     primer_bind     576..617
                     /label=G4mT-pIRES-F
     CDS             597..1508
                     /codon_start=1
                     /label=G4PID
                     /translation="MRSATMHPGHLKGREIGMWYAKKQGQKNKGTGSGAGTGSGAGTGS
                     GAIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGDACIAEYQQAGRGSRG
                     RKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLGADKVRVKWPNDLYLQ
                     DRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWITLQEAGINLDRNTLA
                     AMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGDKEIFGISRGIDKQGAL
                     LLEQDGVIKPWMGGEISLRSAEK"
     CDS             597..683
                     /codon_start=1
                     /label=RHAU23
                     /translation="MRSATMHPGHLKGREIGMWYAKKQGQKNK"
     CDS             684..737
                     /codon_start=1
                     /label=Linker
                     /translation="GTGSGAGTGSGAGTGSGA"
     primer_bind     complement(721..759)
                     /label=G4P-mTurbo-F
     primer_bind     721..759
                     /label=mTurbo-G4P-R
     CDS             738..1508
                     /codon_start=1
                     /product="N-terminally truncated mutant of the E. coli 
                     biotin ligase BioID engineered for improved catalytic 
                     efficiency (Branon et al., 2018)"
                     /label=miniTurbo
                     /translation="IPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGDACI
                     AEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLGADK
                     VRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWITLQ
                     EAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGDKEIF
                     GISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     primer_bind     complement(1492..1532)
                     /label=G4mT-pIRES-R
     polyA_signal    1658..1779
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    1758..1784
                     /label=primer2
     rep_origin      complement(1786..2241)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     misc_feature    1894..1913
                     /label=Feature 18
     promoter        2268..2372
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2374..2731
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2582..2717
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2766..3560
                     /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    3792..3839
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4168..4756
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    4658..4675
                     /label=Feature 19
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 gctagcatga
      601 gatctgccac catgcatcct ggccatctca aaggccgtga aatcggcatg tggtacgcta
      661 agaaacaggg tcagaaaaac aaaggcacgg gaagcggcgc aggcaccggc agcggtgcgg
      721 gtaccggctc cggcgcaatc ccgctgctga acgctaaaca gattctggga cagctggacg
      781 gcgggagcgt ggcagtcctg cctgtggtcg actccaccaa tcagtacctg ctggatcgaa
      841 tcggcgagct gaagagtggg gatgcttgca ttgcagaata tcagcaggca gggagaggaa
      901 gcagagggag gaaatggttc tctccttttg gagctaacct gtacctgagt atgttttggc
      961 gcctgaagcg gggaccagca gcaatcggcc tgggcccggt catcggaatt gtcatggcag
     1021 aagcgctgcg aaagctggga gcagacaagg tgcgagtcaa atggcccaat gacctgtatc
     1081 tgcaggatag aaagctggca ggcatcctgg tggagctggc cggaataaca ggcgatgctg
     1141 cacagatcgt cattggcgcc gggattaacg tggctatgag gcgcgtggag gaaagcgtgg
     1201 tcaatcaggg ctggatcaca ctgcaggaag cagggattaa cctggacagg aatactctgg
     1261 ccgctatgct gatccgagag ctgcgggcag ccctggaact gttcgagcag gaaggcctgg
     1321 ctccatatct gtcacggtgg gagaagctgg ataacttcat caatagaccc gtgaagctga
     1381 tcattgggga caaagagatt ttcgggatta gccgggggat tgataaacag ggagccctgc
     1441 tgctggaaca ggacggagtt atcaaaccct ggatgggcgg agaaatcagt ctgcggtctg
     1501 ccgaaaagta agaattctgc aggtcgagca tgcatagcgg ccgcgactct agatcataat
     1561 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     1621 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     1681 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     1741 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta
     1801 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     1861 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     1921 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     1981 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     2041 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     2101 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     2161 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     2221 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2281 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2341 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     2401 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2461 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     2521 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     2581 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     2641 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     2701 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     2761 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     2821 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     2881 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     2941 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     3001 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     3061 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     3121 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     3181 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     3241 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     3301 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     3361 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     3421 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     3481 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     3541 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     3601 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     3661 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     3721 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     3781 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     3841 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     3901 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     3961 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     4021 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4081 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4141 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4201 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4261 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     4321 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4381 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4441 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4501 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4561 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4621 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4681 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4741 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4801 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     4861 cgccatgcat
//
