LOCUS       sequence-62481-e        8410 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 8410)
  AUTHORS   Jacobson C, Schnapp B, Banker GA
  TITLE     A change in the selective translocation of the Kinesin-1 motor 
            domain marks the initial specification of the axon.
  JOURNAL   Neuron. 2006 Mar 16;49(6):797-804.
  PUBMED    16543128
REFERENCE   2  (bases 1 to 8410)
  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: "Neuron."; 
            date: "2006-03-16"; volume: "49(6)"; pages: "797-804"
COMMENT     Sequence Label: pBa.Kif5c 1-559.GFP
FEATURES             Location/Qualifiers
     source          1..8410
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        418..693
                     /label=chicken beta-actin promoter
     primer_bind     1626..1648
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             2578..2589
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             join(3398..3400,3401..3403,3404..4114)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3398..3400,3401..3403,3404..4114)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3443..3464)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3704..3723)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4051..4072
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4247..4381
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4284..4303)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4338..4357
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4413..4429)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4413..4429)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4426..4448)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4437..4453
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4461..4467,4468..4485,4486..4491))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4506..4527
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4644..4661)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4815..5403)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4895..4914)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5574..6365,6366..6434))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6197..6216
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6435..6539)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6607..6625
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6663..6685)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6785..6804
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6998..7020
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7012..7029
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7013..7029
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ttagtgcatg catacccctc ggtgttattg ctgctctgtg cacgcacgct cattgtatca
       61 cttcatccca gtgcatgcac tcacactgga gcgattgctg ctcggtgcac gcacactcat
      121 tgtatcacgt cagctcagtg gctgcacgca caccggtgtt attgctgctc ggtgcgtgca
      181 tgcacatcag tgtcgctgca gctcagtgca tgcacgctca ttgcccatcg ctatccctgc
      241 ctctcctgct ggcgctcccc gggaggtgac ttcaagggga ccgcaggacc acctcggggg
      301 tggggggagg gctgcacacg cggaccccgc tccccctccc caacaaagca ctgtggaatc
      361 aaaaaggggg gaggggggat ggaggggcgc gtcacacccc cgccccacac cctcacctcg
      421 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt
      481 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggcgc
      541 gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg
      601 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg
      661 cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgttgcc ttcgccccgt
      721 gccccgctcc gcgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca
      781 caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga
      841 cggctcgttt cttttctgtg gctgcgtgaa agccttaaag ggctccggga gggccctttg
      901 tgcggggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg
      961 cggcccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc
     1021 cgcgtgtgcg cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg ggggctgcga
     1081 ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc
     1141 ggcggtcggg ctgtaacccc cccctgcacc cccctccccg agttgctgag cacggcccgg
     1201 cttcgggtgc ggggctccgt gcggggcgtg gcgcggggct cgccgtgccg ggcggggggt
     1261 ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg ggccggggag ggctcggggg
     1321 aggggcgcgg cggccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg
     1381 ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctggcggag
     1441 ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcgggcgaag cggtgcggcg
     1501 ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc
     1561 tccatctcca gcctcggggc tgccgcaggg ggacggctgc cttcgggggg gacggggcag
     1621 ggcggggttc ggcttctggc gtgtgaccgg cggggtttat atcttccctt ctctgttcct
     1681 ccgcagcccc caagctacca tggcagatcc agccgaatgc agcatcaaag tgatgtgccg
     1741 gttccggccc ctcaacgaag ctgagatcct ccgcggggac aaattcatcc ccaaattcaa
     1801 aggcgaggag acggtggtga tcgggcaagg gaagccgtat gtctttgacc gagttctgcc
     1861 gcccaacaca acccaggagc aggtctacaa tgcctgtgcg aagcagattg tcaaagatgt
     1921 cctcgagggt tacaatggga caatttttgc atacgggcag acttcatcag gaaaaactca
     1981 taccatggag gggaagctgc acgatcctca gcttatgggt atcataccca ggatcgcaca
     2041 cgatattttc gatcacatct attccatgga tgagaacttg gagtttcata tcaaggtttc
     2101 ctattttgag atctacttgg acaaaataag ggacttgctt gacgtgtcca agaccaactt
     2161 ggctgttcat gaagacaaaa acagagtccc ctatgtaaag gggtgcaccg agaggtttgt
     2221 gtcaagcccg gaggaggtca tggatgtgat cgatgagggc aaagcaaacc gacatgtggc
     2281 tgtgacaaac atgaacgaac acagctccag gagtcacagt atcttcctga ttaacattaa
     2341 acaagagaat gtggagactg aaaaaaaact cagcgggaag ctgtatttgg ttgatttggc
     2401 tgggagtgaa aaggtcagca aaactggtgc cgagggagct gttcttgatg aagctaaaaa
     2461 tatcaacaag tctttgtctg ctcttggaaa tgtgatctct gccttggcag aagggacaaa
     2521 aacacatgtg ccataccggg acagcaagat gactcggatt ctccaggact ctctgggtgg
     2581 gaactgcagg accaccattg tcatttgctg ttctccttca gtcttcaatg aggctgagac
     2641 caagtccacg ctgatgtttg gacagagagc aaagaccatc aagaatacag tctctgtgaa
     2701 cttggaacta acagcagaag aatggaagaa gaaatatgaa aaagagaaag agaagaataa
     2761 ggccctgaag agtgtcatcc agcatctgga ggtggagctg aacaggtgga ggaacgggga
     2821 agctgtgcct gaggatgaac agatcagtgc caaggaccag aagaacctgg agccctgcga
     2881 caacacaccc atcatagaca acatcacacc tgtggttgac ggcatctctg ctgagaaaga
     2941 gaagtatgac gaggaaatca ccagtctgta ccgacagctg gatgacaagg acgatgaaat
     3001 taaccagcag agccagctgg ctgaaaagct gaagcaacag atgttggatc aggacgagct
     3061 tctagcttcc accagaaggg actacgagaa aatccaggag gagctgaccc gcctccagat
     3121 tgaaaacgag gcggctaaag acgaggtgaa ggaggtcctc caggccctgg aggagctggc
     3181 tgtcaattac gaccagaagt cacaggaagt ggaggacaag acccgggcca acgagcaact
     3241 gaccgatgag ctggcccaga aaacgacgac actgacaacc acccagcggg agctgagtca
     3301 gctacaggag cttagcaacc accagaagaa gagggccacg gagatcctga acctgcttct
     3361 caaggatctg ggcgagcttg gcgcgcctag gcctaccatg gtgagcaagg gcgaggagct
     3421 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     3481 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     3541 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     3601 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     3661 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     3721 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     3781 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     3841 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     3901 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     3961 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     4021 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     4081 cgggatcact ctcggcatgg acgagctgta caagcggacc gtctaatcta gataagtaat
     4141 gatcataatc agccatatca catctgtaga ggttttactt gctttaaaaa acctcccaca
     4201 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     4261 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4321 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     4381 ccccgggtac cgagctcgaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     4441 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     4501 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     4561 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     4621 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4681 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4741 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4801 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4861 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4921 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4981 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5041 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5101 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5161 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5221 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     5281 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5341 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5401 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5461 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5521 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5581 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5641 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5701 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5761 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5821 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5881 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5941 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6001 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6061 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6121 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6181 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6241 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6301 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6361 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6421 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6481 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6541 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     6601 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     6661 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     6721 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ttggcttaac
     6781 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     6841 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     6901 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     6961 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     7021 acggccagtg ccaagttggg atctttgcat tggcccacgg ctctcaggat ggggatgctc
     7081 cccttcagca cccggttccc cttggaaact gatggtcctg gctctgtggc atggcagtgg
     7141 cactgtgagg agcccctacc agcagcacac agtgggtttg gcactgccac gctccggatg
     7201 ccgcgctctg atccaacccc ataatcaagg gaacccgaat tgccccatca ttgcccccac
     7261 cacccccatc ctgccgggcc ctcacacccc acgctgcctt gtggtgacat tccccagccc
     7321 aaacccacgg cttcatggct accgcggggc atttcccatt gccgccccat tatcagctct
     7381 gcacacctcc cgctgtaccc atgcctcgtg gctgcccttc tttgacgtat aatcttctaa
     7441 ttaatacccg gccttgtcaa agtggagcac aaacgttaat taattcccca gcaggcaggt
     7501 aattaacagt gtgactccct ttttgctgcg agtggggctg atacagagag atgtggcact
     7561 atggagccca cggggtcctg gcactgggtg cccacggagg tccccatgtg ctgcagtgtc
     7621 accgcctccg aggtgacagt attgtccctg cggtgtccct gcagctcagc tctgtccaca
     7681 gggccacctc cagtttggag gggacacaat gcagccccga tgcaacccat cctcgcagca
     7741 tcccagggac aaagacccca ctgcaagacc gcacacaggg ctgggtcccg ctcccctaat
     7801 atctacagtg cttttgcatg gccccttaat caatgcagtt aatcagcatg cgctcatgca
     7861 ccgctctgga gctgcaaagc ccctcgcagc gctgctcacc aacaccgcgc accgccccgg
     7921 cccagcctgc agcacgcgct gcaaacagga aagaaacaaa atattgccca aatgtaggca
     7981 aaggcattcg gctgccttga cctccgccgg gccgggccct gcctgactca gctccttact
     8041 cagcgctcgc ttcctccctc cggctgccac cgccgcagcg cacaccctga caaagagtgg
     8101 cccttaacgg gctctgaggt gcacccagca gtgcactcag cagtccaagg gccggcctgg
     8161 aggtttgcac cgctacgtgc tgacattagc attgaacttg gccctgggta gtgctgcagg
     8221 ccgggcgggg tgggtgtaga gagtgcagcg cgcgttgcac ccggtgcccc ttcccctccc
     8281 ttgcatccca gcaggctgca ccccagcacc aggcccgtgc atgcatgctc ctggtgttat
     8341 tgcagcctgg tgcatgcatg cgtcttagtg gtgcagcgct gtgcatgcat cctccttggt
     8401 gtgtagcagc
//
