LOCUS       sequence-495746-        9042 bp    DNA     circular SYN 18-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9042)
  AUTHORS   Burack MA, Silverman MA, Banker G
  TITLE     The role of selective transport in neuronal protein sorting.
  JOURNAL   Neuron. 2000 May;26(2):465-72.
  PUBMED    10839364
REFERENCE   2  (bases 1 to 9042)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 18, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Neuron."; 
            date: "2000-05"; volume: "26(2)"; pages: "465-72"
COMMENT     Sequence Label: pBa.TfR.GFP
FEATURES             Location/Qualifiers
     source          1..9042
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        418..693
                     /label=chicken beta-actin promoter
     primer_bind     1636..1658
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             complement(2667..2678)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      4023..4032
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4029..4031,4032..4034,4035..4745)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4029..4031,4032..4034,4035..4745)
                     /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(4074..4095)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4335..4354)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4682..4703
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4878..5012
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4915..4934)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4969..4988
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5043..5059)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5043..5059)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5056..5078)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5067..5083
                     /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(5091..5097,5098..5115,5116..5121))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5136..5157
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5274..5291)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5445..6033)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5525..5544)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6204..6995,6996..7064))
                     /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     6827..6846
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7065..7169)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7237..7255
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7293..7315)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7415..7434
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7628..7650
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7642..7659
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7643..7659
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ctagtgcatg catacccctc ggtgttattg ctgctctgtg cacgcacgct cattgtatca
       61 cttcatccca gtgcatgcac tcacactgga gcgattgctg ctcggtgcac gcacactcat
      121 tgtatcactg 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 ctgcgcgctg ccttcgcccc
      721 gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc
      781 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa
      841 tgacggctcg tttcttttct gtggctgcgt gaaagcctta aagggctccg ggagggccct
      901 ttgtgcgggg gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc
      961 gtgcggctcc gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg ggctttgtgc
     1021 gctccgcagt gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt gcgggggggg
     1081 ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt
     1141 gggcgcggcg gtcgggctgt aacccccccc tgcacccccc tccccgagtt gctgagcacg
     1201 gcccggcttc gggtgcgggg ctccgtgcgg ggcgtggcgc ggggctcgcc gtgccgggcg
     1261 gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc ggggagggct
     1321 cgggggaggg gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca
     1381 gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct
     1441 gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc
     1501 ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc
     1561 gtccccttct ccatctccag cctcggggct gtccgcaggg ggacggctgc cttcgggggg
     1621 gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggggtttat atcttccctt
     1681 ctctgttcct ccgcagcccc caagcttggc gcgcctaggg gggatccacc atggatcaag
     1741 ctagatcagc attctctaac ttgtttggtg gagaaccatt gtcatatacc cggttcagcc
     1801 tggctcggca agtagatggc gataacagtc atgtggagat gaaacttgct gtagatgaag
     1861 aagaaaatgc tgacaataac acaaaggcca atgtcacaaa accaaaaagg tgtagtggaa
     1921 gtatctgcta tgggactatt gctgtgatcg tctttttctt gattggattt atgattggct
     1981 acttgggcta ttgtaaaggg gtagaaccaa aaactgagtg tgagagactg gcaggaaccg
     2041 agtctccagt gagggaggag ccaggagagg acttccctgc agcacgtcgc ttatattggg
     2101 atgacctgaa gagaaagttg tcggagaaac tggacagcac agacttcacc agcaccatca
     2161 agctgctgaa tgaaaattca tatgtccctc gtgaggctgg atctcaaaaa gatgaaaatc
     2221 ttgcgttgta tgttgaaaat caatttcgtg aatttaaact cagcaaagtc tggcgtgatc
     2281 aacattttgt taagattcag gtcaaagaca gcgctcaaaa ctcggtgatc atagttgata
     2341 agaacggtag acttgtttac ctggtggaga atcctggggg ttatgtggcg tatagtaagg
     2401 ctgcaacagt tactggtaaa ctggtccatg ctaattttgg tactaaaaaa gattttgagg
     2461 atttatacac tcctgtgaat ggatctatag tgattgtcag agcagggaaa atcacctttg
     2521 cagaaaaggt tgcaaatgct gaaagcttaa atgcaattgg tgtgttgata tacatggacc
     2581 agactaaatt tcccattgtt aacgcagaac tttcattctt tggacatgct catctgggga
     2641 caggtgaccc ttacacacct ggattccctt ccttcaatca cactcagttt ccaccatctc
     2701 ggtcatcagg attgcctaat atacctgtcc agacaatctc cagagctgct gcagaaaagc
     2761 tgtttgggaa tatggaagga gactgtccct ctgactggaa aacagactct acatgtagga
     2821 tggtaacctc agaaagcaag aatgtgaagc tcactgtgag caatgtgctg aaagagataa
     2881 aaattcttaa catctttgga gttattaaag gctttgtaga accagatcac tatgttgtag
     2941 ttggggccca gagagatgca tggggccctg gagctgcaaa atccggtgta ggcacagctc
     3001 tcctattgaa acttgcccag atgttctcag atatggtctt aaaagatggg tttcagccca
     3061 gcagaagcat tatctttgcc agttggagtg ctggagactt tggatcggtt ggtgccactg
     3121 aatggctaga gggatacctt tcgtccctgc atttaaaggc tttcacttat attaatctgg
     3181 ataaagcggt tcttggtacc agcaacttca aggtttctgc cagcccactg ttgtatacgc
     3241 ttattgagaa aacaatgcaa aatgtgaagc atccggttac tgggcaattt ctatatcagg
     3301 acagcaactg ggccagcaaa gttgagaaac tcactttaga caatgctgct ttccctttcc
     3361 ttgcatattc tggaatccca gcagtttctt tctgtttttg cgaggacaca gattatcctt
     3421 atttgggtac caccatggac acctataagg aactgattga gaggattcct gagttgaaca
     3481 aagtggcacg agcagctgca gaggtcgctg gtcagttcgt gattaaacta acccatgatg
     3541 ttgaattgaa cctggactat gagaggtaca acagccaact gctttcattt gtgagggatc
     3601 tgaaccaata cagagcagac ataaaggaaa tgggcctgag tttacagtgg ctgtattctg
     3661 ctcgtggaga cttcttccgt gctacttcca gactaacaac agatttcggg aatgctgaga
     3721 aaacagacag atttgtcatg aagaaactca atgatcgtgt catgagagtg gagtatcact
     3781 tcctctctcc ctacgtatct ccaaaagagt ctcctttccg acatgtcttc tggggctccg
     3841 gctctcacac gctgccagct ttactggaga acttgaaact gcgtaaacaa aataacggtg
     3901 cttttaatga aacgctgttc agaaaccagt tggctctagc tacttggact attcagggag
     3961 ctgcaaatgc cctctctggt gacgtttggg acattgacaa tgagtttggg gatccaccgg
     4021 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     4081 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     4141 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     4201 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     4261 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     4321 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     4381 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     4441 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     4501 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     4561 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     4621 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     4681 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     4741 acaagcggac cgtctaatct agataagtaa tgatcataat cagccatatc acatctgtag
     4801 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     4861 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4921 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4981 aactcatcaa tgtatcttat catgtctgga tccccgggta ccgagctcga attcgtaatc
     5041 atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5101 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5161 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5221 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5281 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5341 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5401 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5461 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5521 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5581 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5641 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5701 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5761 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5821 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5881 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5941 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6001 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6061 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6121 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6181 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6241 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6301 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6361 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6421 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6481 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6541 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6601 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6661 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6721 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6781 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6841 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6901 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6961 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7021 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7081 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7141 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     7201 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     7261 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     7321 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     7381 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     7441 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     7501 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     7561 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     7621 gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagttggg atctttgcat
     7681 tggcccacgg ctctcaggat ggggatgctc cccttcagca cccggttccc cttggaaact
     7741 gatggtcctg gctctgtggc atggcagtgg cactgtgagg agccccctac cagcagcaca
     7801 cagtggggtt ggcactgcca cgctccggat gccgcgctct gatccaaccc cataatcaag
     7861 ggaacccgaa ttgccccatc attgccccca ccacccccat cctgccgggc cctcacaccc
     7921 cacgctgcct tctggtgaca ttccccagcc caaacccacg gcttcatggc taccgcgggg
     7981 catttcccat tgccgcccca ttatcagctc tgcacacctc ccgctgtacc catgcctcgt
     8041 ggctgccctt ctttgacgta taatcttcta attaataccc ggccttgtca aagtggagca
     8101 caaacgttaa ttaattcccc agcaggcagg taattaacag tgtgactccc tttttgctgc
     8161 gagtggggct gatacagaga gatgtggcac tatggagccc acggggtcct ggcactgggt
     8221 gcccacggag gtccccatgt gctgcagtgt caccgcctcc gaggagacag tattgtccct
     8281 gcggtgtccc tgcagctcag ctctgtccac agggccacct ccagtttgga ggggacacaa
     8341 tgcagccccg atgcaaccca tcctcgcagc atcccaggga caaagacccc actgcaagac
     8401 cgcacacagg gctgggtccc gctcccctaa tatctacagt gcttcttgca tggcccctta
     8461 atcaatgcag ttaatcagca tgcgctcatg caccgctctg gagctgcaaa gcccctcgca
     8521 gcgctgctca ccaacaccgc gcaccgcccc ggcccagcct gcagcacgcg ctgcaaacag
     8581 gaaagaaaca aaatattgcc caaatgtagg caaaggcatt cggctgcctt gacctccgcc
     8641 gggccgggcc ctgcctgact cagctcctta ctcagcgctc gcttcctccc tccggctgcc
     8701 accgccgcag cgcacaccct gacaaagagt ggcccttaac gggctctgag gtgcacccag
     8761 cagtgcactc agcagtccaa gggccggcct ggaggtttgc accgctacgt gctgacatta
     8821 gcattgaact tggccctggg tagtgctgca ggccgggcgg ggtgggtgta gagagtgcag
     8881 cgcgcgttgc acccggtgcc ccttcccctc ccttgcatcc cagcaggctg caccccagca
     8941 ccaggcccgt gcatgcatgc tcctggtgtt attgcagcct ggtgcatgca tgcgtcttga
     9001 tggtgcagcg ctgtgcatgc atcctccttg gtgtgtagca gt
//
