LOCUS       Exported                4903 bp ds-DNA     circular SYN 20-APR-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4903)
  TITLE     TA Rapoport plasmids
REFERENCE   2  (bases 1 to 4903)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Apr 20, 2021 from SnapGene 5.2.4
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4903
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      614..619
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             620..1480
                     /codon_start=1
                     /label=FLAG-SURF4
                     /translation="MDYKDDDDKSGSGSSAGSGQNDLMGTAEDFADQFLRVTKQYLPHV
                     ARLCLISTFLEDGIRMWFQWSEQRDYIDSTWNCGYLLASSFVFLNLLGQLTGCVLVLSR
                     NFVQYACFGLFGIIALQTIAYSILWDLKFLMRNLALGGGLLLLLAESRSEGKSMFAGVP
                     TMRESSPKQYMQLGGRVLLVLMFMTLLHFDASFFSIVQNIVGTALMILVAIGFKTKLAA
                     LTLVVWLFAINVYFNAFWTIPVYKPMHDFLKYDFFQTMSVIGGLLLVVALGPGGVSMDE
                     KKKEW"
     CDS             623..646
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             674..1480
                     /codon_start=1
                     /label=SURF4
                     /translation="GQNDLMGTAEDFADQFLRVTKQYLPHVARLCLISTFLEDGIRMWF
                     QWSEQRDYIDSTWNCGYLLASSFVFLNLLGQLTGCVLVLSRNFVQYACFGLFGIIALQT
                     IAYSILWDLKFLMRNLALGGGLLLLLAESRSEGKSMFAGVPTMRESSPKQYMQLGGRVL
                     LVLMFMTLLHFDASFFSIVQNIVGTALMILVAIGFKTKLAALTLVVWLFAINVYFNAFW
                     TIPVYKPMHDFLKYDFFQTMSVIGGLLLVVALGPGGVSMDEKKKEW"
     polyA_signal    1698..1819
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1735..1754)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1789..1808
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1826..2281)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1963..1984)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2175..2194
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2308..2412
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2414..2771
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2437..2457)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2622..2757
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2684..2703
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(2707..2718)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2806..3600
                     /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"
     primer_bind     complement(2860..2879)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3470..3489
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3788..3807)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3832..3879
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4208..4796
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4697..4716
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gtcgccacca tggattacaa ggatgacgac gataagtccg gatctggctc
      661 cagcgcaggc agcggccaga acgacctgat gggcacggct gaggacttcg ccgaccagtt
      721 cctccgtgtc acaaagcagt acctgcccca cgtggcgcgc ctctgtctga tcagcacctt
      781 cctggaggac ggcatccgca tgtggttcca gtggagcgag cagcgcgact acatcgacag
      841 cacctggaac tgcggctacc tgctggcctc atccttcgtc ttcctcaact tgctgggaca
      901 gctgactggc tgcgtcctgg tgttgagcag gaacttcgtg cagtacgcct gcttcgggct
      961 ctttggaatc atagctctgc agacgattgc ctacagcatt ttatgggact tgaagttttt
     1021 gatgaggaac ctggccctgg gaggaggcct gttgctgctc ctagcagaat cccgttctga
     1081 agggaagagc atgtttgcgg gcgtccccac catgcgtgag agctccccca aacagtacat
     1141 gcagcttgga ggcagggtct tgctggttct gatgttcatg acccttcttc actttgatgc
     1201 cagcttcttt tctattgtcc agaacatcgt gggcacagct ctgatgattt tagtggccat
     1261 tgggtttaaa accaagctgg ctgctttgac tcttgttgtc tggctctttg ccatcaacgt
     1321 atatttcaac gccttctgga ccattccagt ctacaagccc atgcatgact tcctgaaata
     1381 cgacttcttc cagaccatgt cggtgatcgg gggcttgctc ctggtggtgg cgctgggccc
     1441 tgggggtgtc tccatggatg agaagaagaa ggagtggtaa cagtcacaga tccctacctg
     1501 cctggctaag acccgtggcc gtcgaggact ggtttgggaa ttctgcagtc gacggtaccg
     1561 cgggcccggg atccaccgga tctagataac tgatcataat cagccatacc acatttgtag
     1621 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     1681 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     1741 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     1801 aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     1861 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     1921 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     1981 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     2041 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     2101 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     2161 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     2221 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     2281 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2341 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2401 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     2461 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2521 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2581 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2641 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     2701 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     2761 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     2821 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     2881 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     2941 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     3001 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3061 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3121 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3181 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3241 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3301 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     3361 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     3421 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     3481 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     3541 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     3601 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     3661 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     3721 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     3781 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     3841 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     3901 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     3961 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     4021 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     4081 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4141 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4201 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4261 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4321 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4381 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4441 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4501 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4561 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4621 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4681 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4741 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4801 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4861 ttcctgcgtt atcccctgat tctgtggata accgtattac cgc
//
