LOCUS       sequence-271046-        8613 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 8613)
  AUTHORS   Hussain NK, Jenna S, Glogauer M, Quinn CC, Wasiak S, Guipponi M, 
            Antonarakis SE, Kay BK, Stossel TP, Lamarche-Vane N, McPherson PS
  TITLE     Endocytic protein intersectin-l regulates actin assembly via Cdc42 
            and N-WASP.
  JOURNAL   Nat Cell Biol. 2001 Oct;3(10):927-32.
  PUBMED    11584276
REFERENCE   2  (bases 1 to 8613)
  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: "Nat Cell 
            Biol."; date: "2001-10"; volume: "3(10)"; pages: "927-32"
COMMENT     Sequence Label: GFP-Intersectin Short
FEATURES             Location/Qualifiers
     source          1..8613
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      623..632
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(629..631,632..634,635..1345)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(629..631,632..634,635..1345)
                     /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(674..695)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(935..954)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1282..1303
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5228..5244)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5229..5245)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     misc_feature    5241..5293
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    5417..5538
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5454..5473)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5508..5527
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5545..6000)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5682..5703)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5894..5913
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6027..6131
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6133..6490
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6133..6324
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6156..6176)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6341..6476
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6525..7319
                     /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             6525..7319
                     /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(6579..6598)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7189..7208
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7507..7526)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7551..7598
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7927..8515
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8416..8435
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
      601 agatccgcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
      661 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
      721 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
      781 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
      841 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
      901 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
      961 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     1021 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     1081 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     1141 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     1201 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     1261 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1321 tctcggcatg gacgagctgt acaagtccgg ccggactcag atctcgaggg ctcagtttcc
     1381 aacacctttt ggtggcagcc tggatatctg ggccataact gtagaggaaa gagcgaagca
     1441 tgatcagcag ttccatagtt taaagccaat atctggattc attactggtg atcaagctag
     1501 aaactttttt tttcaatctg ggttacctca acctgtttta gcacagatat gggcactagc
     1561 tgacatgaat aatgatggaa gaatggatca agtggagttt tccatagcta tgaaacttat
     1621 caaactgaag ctacaaggat atcagctacc ctctgcactt ccccctgtca tgaaacagca
     1681 accagttgct atttctagcg caccagcatt tggtatggga ggtatcgcca gcatgccacc
     1741 gcttacagct gttgctccag tgccaatggg atccattcca gttgttggaa tgtctccaac
     1801 cctagtatct tctgttccca cagcagctgt gccccccctg gctaacgggg ctccccctgt
     1861 tatacaacct ctgcctgcat ttgctcatcc tgcagccaca ttgccaaaga gttcttcctt
     1921 tagtagatct ggtccagggt cacaactaaa cactaaatta caaaaggcac agtcatttga
     1981 tgtggccagt gtcccaccag tggcagagtg ggctgttcct cagtcatcaa gactgaaata
     2041 caggcaatta ttcaatagtc atgacaaaac tatgagtgga cacttaacag gtccccaagc
     2101 aagaactatt cttatgcagt caagtttacc acaggctcag ctggcttcaa tatggaatct
     2161 ttctgacatt gatcaagatg gaaaacttac agcagaggaa tttatcctgg caatgcacct
     2221 cattgatgta gctatgtctg gccaaccact gccacctgtc ctgcctccag aatacattcc
     2281 accttctttt agaagagttc gatctggcag tggtatatct gtcataagct caacatctgt
     2341 agatcagagg ctaccagagg aaccagtttt agaagatgag caacaacaat tagaaaagaa
     2401 attacctgta acgtttgaag ataagaagcg ggagaacttt gaacgtggca acctggaact
     2461 ggagaaacga aggcaagctc tcctggaaca gcagcgcaag gagcaggagc gcctggccca
     2521 gctggagcgg gcggagcagg agaggaagga gcgtgagcgc caggagcaag agcgcaaaag
     2581 acaactggaa ctggagaagc aactggaaaa gcagcgggag ctagaacggc agagagagga
     2641 ggagaggagg aaagaaattg agaggcgaga ggctgcaaaa cgggaacttg aaaggcaacg
     2701 acaacttgag tgggaacgga atcgaaggca agaactacta aatcaaagaa acaaagaaca
     2761 agaggacata gttgtactga aagcaaagaa aaagactttg gaatttgaat tagaagctct
     2821 aaatgataaa aagcatcaac tagaagggaa acttcaagat atcagatgtc gattgaccac
     2881 ccaaaggcaa gaaattgaga gcacaaacaa atctagagag ttgagaattg ccgaaatcac
     2941 ccatctacag caacaattac aggaatctca gcaaatgctt ggaagactta ttccagaaaa
     3001 acagatactc aatgaccaat taaaacaagt tcagcagaac agtttgcaca gagattcact
     3061 tgttacactt aaaagagcct tagaagcaaa agaactagct cggcagcacc tacgagacca
     3121 actggatgaa gtggagaaag aaactagatc aaaactacag gagattgata ttttcaataa
     3181 tcagctgaag gaactaagag aaatacacaa taagcaacaa ctccagaagc aaaagtccat
     3241 ggaggctgaa cgactgaaac agaaagaaca agaacgaaag atcatagaat tagaaaaaca
     3301 aaaagaagaa gcccaaagac gagctcagga aagggacaag cagtggctgg agcatgtgca
     3361 gcaggaggac gagcatcaga gaccaagaaa actccacgaa gaggaaaaac tgaaaaggga
     3421 ggagagtgtc aaaaagaagg atggcgagga aaaaggcaaa caggaagcac aagacaagct
     3481 gggtcggctt ttccatcaac accaagaacc agctaagcca gctgtccagg caccctggtc
     3541 cactgcagaa aaaggtccac ttaccatttc tgcacaggaa aatgtaaaag tggtgtatta
     3601 ccgggcactg tacccctttg aatccagaag ccatgatgaa atcactatcc agccaggaga
     3661 catagtcatg gttaaagggg aatgggtgga tgaaagccaa actggagaac ccggctggct
     3721 tggaggagaa ttaaaaggaa agacagggtg gttccctgca aactatgcag agaaaatccc
     3781 agaaaatgag gttcccgctc cagtgaaacc agtgactgat tcaacatctg cccctgcccc
     3841 caaactggcc ttgcgtgaga cccccgcccc tttggcagta acctcttcag agccctccac
     3901 gacccctaat aactgggccg acttcagctc cacgtggccc accagcacga atgagaaacc
     3961 agaaacggat aactgggatg catgggcagc ccagccctct ctcaccgttc caagtgccgg
     4021 ccagttaagg cagaggtccg cctttactcc agccacggcc actggctcct ccccgtctcc
     4081 tgtgctaggc cagggtgaaa aggtggaggg gctacaagct caagccctat atccttggag
     4141 agccaaaaaa gacaaccact taaattttaa caaaaatgat gtcatcaccg tcctggaaca
     4201 gcaagacatg tggtggtttg gagaagttca aggtcagaag ggttggttcc ccaagtctta
     4261 cgtgaaactc atttcagggc ccataaggaa gtctacaagc atggattctg gttcttcaga
     4321 gagtcctgct agtctaaagc gagtagcctc tccagcagcc aagccggtcg tttcgggaga
     4381 agaatttatt gccatgtaca cttacgagag ttctgagcaa ggagatttaa cctttcagca
     4441 aggggatgtg attttggtta ccaagaaaga tggtgactgg tggacaggaa cagtgggcga
     4501 caaggccgga gtcttccctt ctaactatgt gaggcttaaa gattcagagg gctctggaac
     4561 tgctgggaaa acagggagtt taggaaaaaa acctgaaatt gcccaggtta ttgcctcata
     4621 caccgccacc ggccccgagc agctcactct cgcccctggt cagctgattt tgatccgaaa
     4681 aaagaaccca ggtggatggt gggaaggaga gctgcaagca cgtgggaaaa agcgccagat
     4741 aggctggttc ccagctaatt atgtaaagct tctaaaccct gggacgagca aaatcactcc
     4801 aacagagcca cctaagtcaa cagcattagc ggcagtgtgc caggtgattg ggatgtacga
     4861 ctacaccgcg cagaatgacg atgagctggc cttcaacaag ggccagatca tcaacgtcct
     4921 caacaaggag gaccctgact ggtggaaagg agaagtcaat ggacaagtgg ggctcttccc
     4981 atccaattat gtgaagctga ccacagacat ggacccaagc cagcaatgaa tcatatgttg
     5041 tccacccccc cctcaggctt gaaagtcctc aaagagaccc actatcccag cttggactta
     5101 accaggctga acttgctagt catggttaaa ggggaatggg tggatgaaag ccaaactgga
     5161 taacccggct ggcttggagg agaattaaaa ggaaagacag ggtgccggaa ttcgatatca
     5221 agcttatcga taccgtcgac ctcgagctca agcttcgaat tctgcagtcg acggtaccgc
     5281 gggcccggga tccaccggat ctagataact gatcataatc agccatacca catttgtaga
     5341 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     5401 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5461 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5521 actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     5581 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     5641 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     5701 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     5761 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     5821 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     5881 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     5941 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     6001 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     6061 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     6121 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     6181 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     6241 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     6301 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     6361 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     6421 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     6481 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     6541 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     6601 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     6661 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     6721 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     6781 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     6841 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     6901 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     6961 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     7021 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     7081 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     7141 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     7201 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     7261 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     7321 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     7381 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     7441 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     7501 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     7561 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     7621 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     7681 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     7741 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     7801 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     7861 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     7921 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     7981 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     8041 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     8101 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     8161 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     8221 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     8281 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     8341 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     8401 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     8461 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     8521 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     8581 tcctgcgtta tcccctgatt ctgtggataa ccg
//
