LOCUS       sequence-14492-e        7504 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 7504)
  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 7504)
  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: pKin1A
FEATURES             Location/Qualifiers
     source          1..7504
                     /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    4292..4413
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4329..4348)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4383..4402
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4420..4875)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4557..4578)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4769..4788
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4902..5006
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5008..5365
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5008..5199
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5031..5051)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5216..5351
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5400..6194
                     /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             5400..6194
                     /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(5454..5473)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6064..6083
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6382..6401)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6426..6473
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6802..7390
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7291..7310
                     /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 ccggagtaca cgctaccctt gcagtcggag acgctcccag cccctcgtcg tcgtcgacca
      661 ccatggcgga gactaacaac gaatgcagca tcaaggtgct ttgccgattt cggcccctga
      721 accaggccga gattctgcgg ggggacaagt tcatccccat tttccaaggg gacgacagcg
      781 tcattattgg gggaaagcca tatgtctttg accgcgtctt ccccccaaac accactcagg
      841 agcaggttta ccacgcctgt gccatgcaga tcgtcaaaga cgtccttgct ggttacaatg
      901 gcacaatctt cgcttatgga cagacatcct cagggaaaac gcataccatg gaggggaagc
      961 tgcacgaccc tcagctgatg ggcatcattc cccggatcgc tcgagacatc ttcaaccaca
     1021 tctactccat ggatgagaac cttgaattcc acattaaggt atcttacttc gagatttacc
     1081 tggataagat ccgtgacctt ttggatgtga ccaagacgaa cctgtccgtg catgaggaca
     1141 aaaaccgggt gccgtttgtc aagggttgta ccgaacgctt tgtgtccagc ccagaggaga
     1201 ttctggatgt gatcgatgag gggaagtcca accgtcacgt agctgtcacc aacatgaacg
     1261 agcacagttc tcggagccac agcatcttcc tcatcaacat caagcaggag aacgtagaga
     1321 ccgagcagaa gctcagcggg aagctgtacc tcgtggatct ggccggaagc gagaaggtca
     1381 gcaagacagg ggcagaggga gccgttctgg acgaggcaaa gaatatcaac aagtcgctgt
     1441 cggccctggg gaacgtgatc tctgcactgg cagagggcac caaaagctac gtgccgtacc
     1501 gcgacagcaa aatgacgagg attctccagg actctctggg agggaactgc aggactacca
     1561 tgttcatctg ctgctcgccg tccagctaca atgacgcaga gaccaagtcc acgctcatgt
     1621 ttggacagcg ggcgaagacc atcaagaaca ctgcctcagt gaatctggag ctgactgctg
     1681 agcagtggaa gaagaagtat gagaaggaga aggagaagac caaggcccag aaggagacaa
     1741 ttgcgaagct agaggctgag cttagccggt ggcgcaatgg agagaatgtg cctgagactg
     1801 agcgcctggc tggagaggac tcagctctgg gagctgagct ctgcgaggag acccctgtga
     1861 atgacaactc atccattgtg gtacgcatcg cacctgagga aaggcagaaa tatgaggaag
     1921 agatccgccg tctctacaag cagcttgatg acaaggatga tgagatcaac cagcagagcc
     1981 agctcattga gaagctgaag cagcagatgc tggaccagga agagctgctc gtgtccactc
     2041 ggggagacaa cgagaaggtc cagcgggagc ttagccacct gcagtccgag aacgatgctg
     2101 cgaaggacga ggtgaaggaa gtgctgcagg ccctagagga gctggcggtc aactacgacc
     2161 agaagtccca ggaggtggag gagaagagcc agcagaacca gctgctggtg gacgagctgt
     2221 cccagaaagt ggccaccatg ctgtccctgg agtccgagct acagcggctc caggaggtca
     2281 gtggacacca gcgaaagcgg atcgctgagg tgctgaatgg gctgatgagg gacctgagtg
     2341 agttcagtgt catcgtgggc aacggcgaga ttaagctgcc cgtggagatc agtggggcca
     2401 tcgaggagga gttcacggtg gcccggctct acatcagcaa gatcaagtcg gaggtgaagt
     2461 ccgtggttaa gcgatgtcgg cagctggaga acctccaggt ggagtgtcat cgcaagatgg
     2521 aggtgaccgg tagggagctg tcatcttgcc aactgctcat ctcacagcat gaggccaaga
     2581 tccgttcact cacggagtac atgcagactg tggagttgaa gaaacggcac ctggaagagt
     2641 cctacgactc cctgagcgat gagcttgcca ggctccaggc gcacgaaact gtacacgagg
     2701 tagctctgaa agacaaggag ccagacacac aggacgcgga ggaggtgaag aaggccctgg
     2761 aactacagat ggagaatcac cgtgaggccc atcaccggca gctggcccgc ctccgggatg
     2821 agattaatga gaaacagaaa accattgatg agctgaaaga cctgaaccag aagctccagt
     2881 tagagctgga gaagcttcag gccgactatg agaggctgaa gaatgaagag aacgagaaga
     2941 gcgccaagct ccaggagctg acatttctgt atgagcgaca tgagcagtcc aagcaggacc
     3001 tcaaggggct ggaggagaca gttgcccgtg aactccagac cctccacaac cttcgcaagc
     3061 tgttcgttca agacgtcacg actcgagtca agaaaagtgc agaaatggag cccgaggaca
     3121 gtggggggat tcattcccaa aagcagaaga tctcctttct tgagaacaac ctggaacagc
     3181 ttacaaaggt tcacaaacag ctggtacgtg acaatgcaga tctgcgttgt gagcttccta
     3241 aattggaaaa acgacttcgg gctacggctg agagagttaa ggccctggag ggtgcactga
     3301 aggaggccaa ggagggcgct atgaaggaca agcgtagata ccagcaggag gtggaccgca
     3361 tcaaagaagc cgtgcggtac aagagctccg gcaagcgggg ccattctgcc cagatcgcta
     3421 agcctgtgag gcctggccac tatcctgcct cctcacccac caacccctac ggcacccgga
     3481 gccccgagtg tatcagctac accaacaacc tcttccagaa ctaccagaac ctgcacctgc
     3541 aggctgcgcc tagctccact tcagatatgt actttgccag cagcggagcc acatctgttg
     3601 cccccttggc ttcctaccag aaggccaaca tggacaatgg aaatgccaca gatatcaacg
     3661 acaacaggag tgacctgccg tgtggctatg aggctgagga ccaggccaag cttttccctc
     3721 tccaccaaga gacagcagcc agctaactcc caaccctggc cacatatctg cactttcagg
     3781 tagctgcagg ccacttccct ccctagcccc agcctcctcc tgtctggcca tggctgcttc
     3841 tcctgttcat gtgtatgtgt atatgtttgt atgtgtctgt atgtgtgtgt tgtgtgtgca
     3901 tgagtgtacc cacatcagga ggccaaggtt agtaggtgtg ggtgtcttca tctgtgtgtg
     3961 tgtgtgtgtg cgcgctcaca tccacaggcc agccttggtg tgggtgtctt cctctctcac
     4021 tttccatctt actttttgag gtctctcaca gaacctggag cttgctgttt tggggtagac
     4081 tggctggtca gtcacctccc caccctccct ctgtcccacc ccacccccca tatctgccca
     4141 tttctgtccc ctggggtact ggggatccac cggatctaga taactgatca taatcagcca
     4201 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     4261 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     4321 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4381 ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt
     4441 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     4501 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     4561 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     4621 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     4681 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     4741 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     4801 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     4861 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     4921 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     4981 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     5041 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     5101 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5161 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     5221 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     5281 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     5341 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     5401 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     5461 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     5521 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     5581 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     5641 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     5701 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     5761 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     5821 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     5881 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     5941 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     6001 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     6061 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     6121 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     6181 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     6241 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     6301 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     6361 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     6421 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     6481 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     6541 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     6601 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     6661 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6721 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     6781 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6841 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6901 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     6961 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     7021 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     7081 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     7141 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     7201 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     7261 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     7321 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     7381 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7441 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
     7501 gcat
//
