LOCUS       sequence-159443-        6498 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6498)
  AUTHORS   Joseph J, Tan SH, Karpova TS, McNally JG, Dasso M
  TITLE     SUMO-1 targets RanGAP1 to kinetochores and mitotic spindles.
  JOURNAL   J Cell Biol. 2002 Feb 18. 156(4):595-602.
  PUBMED    11854305
REFERENCE   2  (bases 1 to 6498)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell 
            Biol. 2002 Feb 18. 156(4):595-602."
COMMENT     Sequence Label: pDsRed1-N1 RanGAP
FEATURES             Location/Qualifiers
     source          1..6498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        16..319
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        320..523
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     474..494
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1539..1556
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     regulatory      2434..2443
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2440..2442,2443..2445,2446..3120)
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLFL"
     primer_bind     complement(2623..2643)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2841..2860
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3035..3058
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    3241..3362
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3278..3297)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3332..3351
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3369..3824)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3506..3527)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3718..3737
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3851..3955
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3957..4314
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3957..4148
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3980..4000)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4165..4300
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4349..5143
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4349..5143
                     /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(4403..4422)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5013..5032
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5331..5350)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5375..5422
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5751..6339
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6240..6259
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
       61 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      301 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      541 gatccgctag cgctaccgga ctcagatctc gagctcaagc ttcgaattct gcagtcgacg
      601 gtaccgcggg ccgggatggc ctcggaagac attgccaagc tggcagagac acttgccaag
      661 actcaggtgg ccgggggaca gctgagtttc aaaggcaaga gcctcaaact caacactgca
      721 gaagatgcta aagatgtgat taaagagatt gaagactttg acagcttgga ggctctgcgt
      781 ctggaaggca acacagtggg cgtggaagcc gccagggtca tcgccaaggc cttagagaag
      841 aagtcggagt tgaagcgctg ccactggagt gacatgttca cgggaaggct gcggaccgag
      901 atcccaccag ccctgatctc actaggggaa ggactcatca cagctggggc tcagctggtg
      961 gagctggact taagcgacaa cgcattcggg cccgacggtg tgcaaggctt cgaggccctg
     1021 ctcaagagct cagcctgctt caccctgcag gaactcaagc tcaacaactg tggcatgggc
     1081 attggcggcg gcaagatcct ggctgcagct ctgaccgaat gtcaccggaa atccagtgcc
     1141 caaggcaagc ctctggccct gaaggtcttt gtggctggca gaaaccgtct ggagaatgat
     1201 ggcgccactg ccttggcaga agcttttagg gtcatcggga ccctggagga ggtccacatg
     1261 ccacagaatg ggatcaacca ccctggcatc actgccctgg cccaggcttt cgctgtcaac
     1321 cccctgctgc gggtcatcaa cctgaatgac aacaccttca ctgagaaggg cgccgtggcc
     1381 atggccgaga ccttgaagac cttgcggcag gtggaggtga ttaattttgg ggactgcctg
     1441 gtgcgctcca agggtgcagt tgccattgca gatgccatcc gcggcggcct gcccaagcta
     1501 aaggagctga acttgtcatt ctgtgagatc aagagggatg ctgccctggc tgttgctgag
     1561 gccatggcag acaaagctga gctggagaag ctggacctga atggcaacac cctgggagaa
     1621 gaaggctgtg aacagcttca ggaggtgctg gagggcttca acatggccaa ggtgctggcg
     1681 tccctcagtg atgacgagga cgaggaggag gaggaggaag gagaagagga agaagaggaa
     1741 gcagaagaag aggaggagga agatgaggaa gaggaggaag aagaggagga ggaggaggaa
     1801 gaagagcctc agcagcgagg gcagggagag aagtcagcca cgccctcacg gaagattctg
     1861 gaccctaaca ctggggagcc agctcccgtg ctgtcctccc cacctcctgc agacgtctcc
     1921 accttcctgg cttttccctc tccagagaag ctgctgcgcc tagggcccaa gagctccgtg
     1981 ctgatagccc agcagactga cacgtctgac cccgagaagg tggtctctgc cttcctaaag
     2041 gtgtcatctg tgttcaagga cgaagctact gtgaggatgg cagtgcagga tgcagtagat
     2101 gccctgatgc agaaggcttt caactcctcg tccttcaact ccaacacctt cctcaccagg
     2161 ctcctcgtgc acatgggtct gctcaagagt gaagacaagg tcaaggccat tgccaacctg
     2221 tacggccccc tgatggcgct gaaccacatg gtgcagcagg actatttccc caaggccctt
     2281 gcacccctgc tgctggcgtt cgtgaccaag cccaacagcg ccctggaatc ctgctccttc
     2341 gcccgccaca gtctgctgca gacgctgtac aaggtcaagg gcaattctgc agatatccag
     2401 cacagtggcg gccgctcgag ggatccaccg gtcgccacca tggtgcgctc ctccaagaac
     2461 gtcatcaagg agttcatgcg cttcaaggtg cgcatggagg gcaccgtgaa cggccacgag
     2521 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gccacaacac cgtgaagctg
     2581 aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc ccagttccag
     2641 tacggctcca aggtgtacgt gaagcacccc gccgacatcc ccgactacaa gaagctgtcc
     2701 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
     2761 gtgacccagg actcctccct gcaggacggc tgcttcatct acaaggtgaa gttcatcggc
     2821 gtgaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     2881 accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg agatccacaa ggccctgaag
     2941 ctgaaggacg gcggccacta cctggtggag ttcaagtcca tctacatggc caagaagccc
     3001 gtgcagctgc ccggctacta ctacgtggac tccaagctgg acatcacctc ccacaacgag
     3061 gactacacca tcgtggagca gtacgagcgc accgagggcc gccaccacct gttcctgtag
     3121 cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta
     3181 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     3241 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     3301 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     3361 taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     3421 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     3481 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     3541 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     3601 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     3661 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     3721 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     3781 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     3841 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3901 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     3961 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4021 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4081 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4141 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4201 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     4261 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     4321 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     4381 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4441 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4501 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     4561 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4621 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4681 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4741 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4801 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4861 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4921 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4981 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     5041 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     5101 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     5161 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     5221 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     5281 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     5341 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     5401 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     5461 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     5521 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     5581 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     5641 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5701 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5761 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5821 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5881 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5941 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6001 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6061 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6121 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6181 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6241 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6301 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6361 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6421 gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat caattacggg
     6481 gtcattagtt catagccc
//
