LOCUS       sequence-12460-e        6189 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 6189)
  AUTHORS   Thyagarajan A, Ting AY
  TITLE     Imaging activity-dependent regulation of neurexin-neuroligin 
            interactions using trans-synaptic enzymatic biotinylation.
  JOURNAL   Cell. 2010 Oct 29;143(3):456-69. doi: 10.1016/j.cell.2010.09.025. 
            Epub 2010 Oct 7.
  PUBMED    20933261
REFERENCE   2  (bases 1 to 6189)
  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"; doi: 
            "10.1016/j.cell.2010.09.025"; journalName: "Cell"; date: 
            "2010-10-29- 29"; volume: "143"; issue: "3"; pages: "456-69"
COMMENT     Sequence Label: p-ECFP-N1-BirA-neurexin
FEATURES             Location/Qualifiers
     source          1..6189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        285..488
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     439..459
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             746..775
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             776..1735
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAIGLSLVIGIVMAEVLRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     polyA_signal    2977..3098
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3014..3033)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3068..3087
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3105..3560)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3242..3263)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3454..3473
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3587..3691
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3693..4050
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3693..3884
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3716..3736)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3901..4036
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4085..4879
                     /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             4085..4879
                     /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(4139..4158)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4749..4768
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5067..5086)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5111..5158
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5487..6075
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5976..5995
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
       61 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      121 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      181 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      241 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      301 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      361 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
      421 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
      481 gcagagctgg tttagtgaac cgtcagatcc gctagcgcta ccggactcag atctcgagct
      541 caagcttacc atgtaccaga ggatgctccg gtgcggcgcc gatctgggat cgcccggggg
      601 cggcagtggc ggcggcgcag gggggcgcct ggccctgatc tggatagtcc cgctcaccct
      661 cggcggcctc ctaggagtgg cctggggggc atccagtttg ggagcgcacc acatccacca
      721 tttccatggc agcagcaagg aattcgaaca aaaactcatc tcagaagagg atctaatgaa
      781 ggataacacc gtgccactga aattgattgc cctgttagcg aacggtgaat ttcactctgg
      841 cgagcagttg ggtgaaacgc tgggaatgag ccgggcggct attaataaac acattcagac
      901 actgcgtgac tggggcgttg atgtctttac cgttccgggt aaaggataca gcctgcctga
      961 gcctatccag ttacttaatg ctaaacagat attgggtcag ctggatggcg gtagtgtagc
     1021 cgtgctgcca gtgattgact ccacgaatca gtaccttctt gatcgtatcg gagagcttaa
     1081 atcgggcgat gcttgcattg cagaatacca gcaggctggc cgtggtcgcc ggggtcggaa
     1141 atggttttcg ccttttggcg caaacttata tttgtcgatg ttctggcgtc tggaacaagg
     1201 cccggcggcg attggtttaa gtctggttat cggtatcgtg atggcggaag tattacgcaa
     1261 gctgggtgca gataaagttc gtgttaaatg gcctaatgac ctctatctgc aggatcgcaa
     1321 gctggcaggc attctggtgg agctgactgg caaaactggc gatgcggcgc aaatagtcat
     1381 tggagccggg atcaacatgg caatgcgccg tgttgaagag agtgtcgtta atcaggggtg
     1441 gatcacgctg caggaagcgg ggatcaatct cgatcgtaat acgttggcgg ccatgctaat
     1501 acgtgaatta cgtgctgcgt tggaactctt cgaacaagaa ggattggcac cttatctgtc
     1561 gcgctgggaa aagctggata attttattaa tcgcccagtg aaacttatca ttggtgataa
     1621 agaaatattt ggcatttcac gcggaataga caaacagggg gctttattac ttgagcagga
     1681 tggaataata aaaccctgga tgggcggtga aatatccctg cgtagtgcag aaaaaactag
     1741 tcctattgca atctacaggt caccagcatc cttgcgaggc ggacacgctg ggacaacata
     1801 tatctttagc aaaggtggtg gacagattac atataagtgg cctcccaatg accgccccag
     1861 tacacgagca gacaggctgg ccatcggatt tagcactgtc cagaaggaag ccgtgttggt
     1921 gcgtgtggac agttcctcag gcctgggtga ctacctggag ctgcacatac accaaggaaa
     1981 aattggagtt aagtttaatg ttgggacaga tgacatcgcc atcgaagagt ctaatgcaat
     2041 cattaatgat gggaaatacc atgtagtacg tttcacaagg agtggtggca atgccacgtt
     2101 acaggtggac agctggccag ttatcgaacg ctaccctgca gggcgtcagc tcacaatctt
     2161 caatagccaa gcaaccataa taattggcgg gaaagagcag ggccagccct tccagggcca
     2221 gctctctggt ctttactaca atggcttgaa agttctgaat atggcagcag agaacgatgc
     2281 caacatcgcc atagtgggga atgtgaggct ggtcggtgaa gtgccttcct ctatgacaac
     2341 tgagtcgaca gccactgcca tgcagtccga gatgtccacc tcaatcatgg agaccaccac
     2401 aaccctggct accagcacag ctcgacgagg aaagcccccc acaaaggagc ctatcagcca
     2461 gaccacggat gatatccttg tggcctcggc agagtgtccc agcgacgatg aggacattga
     2521 cccctgtgag ccgagctcag gtgggttagc caaccccacc cgagtgggcg gccgcgaacc
     2581 atacccaggc tcggcagagg tgatccggga gtctagcagt accactggca tggtggtggg
     2641 gattgtcgca gcagcagctc tgtgcatcct catcctcctc tatgccatgt acaagtacag
     2701 aaaccgggat gaaggctcat atcacgtgga tgagagtcga aactacatca gtaactcagc
     2761 acagtccaat ggggctgtgg tcaaggagaa gcagcccagc agtgcgaaaa gcgccaacaa
     2821 aaacaagaag aacaaggata aggagtatta tgtctaggga tccgactcta gatcataatc
     2881 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     2941 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     3001 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3061 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
     3121 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     3181 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     3241 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     3301 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     3361 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     3421 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     3481 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     3541 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3601 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3661 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     3721 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3781 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3841 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3901 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3961 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     4021 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     4081 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     4141 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     4201 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4261 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4321 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4381 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4441 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4501 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4561 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     4621 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4681 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4741 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4801 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4861 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4921 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4981 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     5041 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     5101 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     5161 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     5221 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     5281 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     5341 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5401 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5461 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5521 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5581 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     5641 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5701 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5761 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5821 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5881 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5941 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6001 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6061 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6121 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6181 gccatgcat
//
