LOCUS       sequence-36093-e        7273 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7273)
  AUTHORS   Uchida A, Alami NH, Brown A
  TITLE     Tight functional coupling of kinesin-1A and dynein motors in the 
            bidirectional transport of neurofilaments.
  JOURNAL   Mol Biol Cell. 2009 Dec . 20(23):4997-5006.
  PUBMED    19812246
REFERENCE   2  (bases 1 to 7273)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Biol 
            Cell. 2009 Dec . 20(23):4997-5006."
COMMENT     Sequence Label: pKin1B
FEATURES             Location/Qualifiers
     source          1..7273
                     /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     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    4061..4182
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4098..4117)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4152..4171
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4189..4644)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4326..4347)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4538..4557
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4671..4775
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4777..5134
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4777..4968
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4800..4820)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4985..5120
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5169..5963
                     /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             5169..5963
                     /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(5223..5242)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5833..5852
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6151..6170)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6195..6242
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6571..7159
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7060..7079
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
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 gctagcgcta
      601 ccggtagtga acttcgggcg caggctgccg gcgacagcgc ttcggtctag ctgcccgggg
      661 tgcacggcgc tgcggagccc ggaccttccg gtcctcgcga cccctgcctc ttggctcccc
      721 tcgcttaccc caagactgag cggcttgaag aggcgcccgg cctgcggtcc cggcttccag
      781 aaagatggcg gacccggcgg agtgcaacat caaagtgatg tgtcgcttca gacctctcaa
      841 cgaatctgaa gtgaaccgcg gcgataagta cgtcgccaaa ttccagggag aagacacggt
      901 gatgatcgcg tccaagcctt atgcctttga tcgtgtgttc cagtcaagca catctcaaga
      961 gcaagtatac aacgactgtg caaaaaagat tgttaaagat gttcttgagg gctataatgg
     1021 aacaatattt gcatatggac aaacatcatc tgggaagacc cacacgatgg agggtaaact
     1081 tcatgatcca gaaggcatgg gaattattcc aagaatagtg caagatattt ttaattatat
     1141 ttactccatg gatgaaaatt tggaatttca tattaaggtt tcatattttg aaatatattt
     1201 ggataagata agggacttgt tagatgtttc aaagactaac ctttcagtcc atgaagacaa
     1261 aaaccgtgtt ccctatgtaa aggggtgcac agaacgtttc gtgtgtagtc cagatgaagt
     1321 catggatacc atagatgaag ggaaatccaa cagacatgtc gcagttacaa atatgaatga
     1381 acatagctct aggagtcaca gcatatttct tattaatgta aaacaagaga atacacaaac
     1441 ggaacagaaa ctcagtggaa agctttatct ggttgattta gctggcagtg agaaggttag
     1501 taagactggg gctgaaggtg ctgtgctgga tgaagctaag aacatcaaca agtcactttc
     1561 tgcacttgga aatgtcattt ctgctttggc agagggcagt acctatgttc cttatcgaga
     1621 tagtaaaatg accagaattc ttcaagattc attaggtggc aactgtagga ccactattgt
     1681 catatgctgc tctccatcat catacaatga gtctgagaca aagtcaacac tcctctttgg
     1741 tcaaagggcc aaaacaatta agaacacagt ctgtgtcaat gtagagttaa ctgcggagca
     1801 gtggaaaaag aagtatgaaa aagaaaagga aaaaaataaa actctacgga acactattca
     1861 gtggctggaa aacgagctaa accgttggcg taacggggag acagtgccta ttgatgagca
     1921 gtttgacaaa gagaaagcta atttggaagc cttcacagcg gataaagata ttgctattac
     1981 cagtgataaa ccagctgctg cagtcggaat ggctggtagt tttaccgatg ctgaaagaag
     2041 aaagtgtgaa gaagaacttg ctaaattgta taaacaactt gatgacaagg atgaagagat
     2101 taaccaacaa agccaattgg tagagaaatt gaagacacaa atgctggatc aggaagagct
     2161 tctggcatca accagaaggg atcaagataa tatgcaagct gaactgaatc gcctccaagc
     2221 agaaaatgat gcttctaaag aagaagtcaa agaagtttta caggccttag aggaactggc
     2281 tgttaattat gatcagaagt ctcaggaagt tgaagacaaa acaaaggaat atgaattgct
     2341 tagtgatgaa ttgaatcaaa aatctgcaac tttagcaagt attgatgctg agcttcagaa
     2401 gctgaaggaa atgaccaacc accagaagaa acgagcagct gaaatgatgg catcattatt
     2461 aaaagacctt gcagaaatag gaattgctgt ggggaataac gatgtgaagc aaccagaagg
     2521 aactggtatg atagatgaag agtttactgt tgcaagactc tacattagca aaatgaaatc
     2581 agaagtaaag accatggtga aacgctgcaa acagctagaa agcacacaga ctgagagcaa
     2641 caaaaaaatg gaagaaaatg agaaagagtt agcagcatgc cagcttcgga tctcccaaca
     2701 tgaagccaaa atcaagtcac tgactgagta ccttcagaat gtagaacaaa agaagaggca
     2761 gctggaggag tctgttgatt cccttggtga ggagctagtc caactccgag cacaagagaa
     2821 agtccatgaa atggaaaaag agcacttgaa caaggttcag actgcaaatg aagtcaagca
     2881 agctgttgag cagcagatcc agagtcacag agaaacccac caaaaacaaa tcagtagttt
     2941 gcgagatgaa gtggaggcaa aggaaaagct aatcactgac ctccaagacc aaaaccagaa
     3001 gatggtgttg gagcaggaac ggctaagggt ggagcatgag aggctgaagg ctacagacca
     3061 agagaagagc aggaagctgc acgagctcac ggttatgcaa gacagacgag aacaagcaag
     3121 acaagacttg aagggtttgg aggagaccgt ggcaaaagaa cttcagactt tacacaacct
     3181 gcgtaagctc tttgttcagg acttggctac cagggtgaaa aagagcgccg aggtcgactc
     3241 tgacgacact ggcggcagtg ctgcacagaa gcagaaaatc tccttccttg aaaacaacct
     3301 tgaacagctc accaaagtgc acaagcagtt ggtacgcgat aatgcagatc ttcgctgtga
     3361 gcttcctaag ttagagaaac ggcttagagc tactgcagaa agagtgaaag ctttggagtc
     3421 agccctgaaa gaagccaaag aaaatgcatc tcgagaccgt aaacgctatc agcaagaagt
     3481 agaccggata aaggaagcag tcaggtcaaa gaacatggcc agaaggggac attctgccca
     3541 gattgcaaag ccgatccgtc ctggacagca tccagcggcc tcgccaactc acccgggcac
     3601 agttcgtgga ggaggctcat ttgttcagaa caaccagcca gtggggcttc gtggtggtgg
     3661 aggcaagcag tcgtaaacac ttgacacatt gcaggtgttg aaaagtaatt gaagtttgaa
     3721 gaggacataa aatcagtctt tcactaacta acctctattc cttgggattg tagaaattta
     3781 attttttaat gtgtaaatta ctcctaccct gtacagctgt ttcctaccca ctcttgtaaa
     3841 ctctgctgct tcccaacact agagtgcaat gttgatgtcc tagaagggaa acgacagttt
     3901 acaactgcgg ccctgtttgt tacacggtgt gggatccacc ggatctagat aactgatcat
     3961 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     4021 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     4081 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4141 gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg
     4201 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     4261 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     4321 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     4381 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     4441 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     4501 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     4561 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     4621 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4681 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4741 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     4801 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4861 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4921 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4981 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     5041 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     5101 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     5161 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     5221 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     5281 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     5341 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     5401 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     5461 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     5521 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     5581 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     5641 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     5701 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     5761 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     5821 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     5881 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     5941 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     6001 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     6061 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     6121 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     6181 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     6241 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     6301 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     6361 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     6421 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     6481 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     6541 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6601 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6661 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     6721 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6781 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6841 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6901 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6961 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     7021 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     7081 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     7141 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     7201 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     7261 taccgccatg cat
//
