LOCUS       sequence-379648-        8229 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Destination Vector, containing NOA1-GFP chimera coding 
            sequence, for Gateway Cloning Technology.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8229)
  AUTHORS   Scorrano L
  TITLE     Luca Scorrano plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 8229)
  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: 
            "unpublished"
COMMENT     Sequence Label: pcDNA-DEST47-Noa1-GFP
FEATURES             Location/Qualifiers
     source          1..8229
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(60..80)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        65..394
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      245..380
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     307..326
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             461..1255
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(515..534)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1125..1144
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1431..1552
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1468..1487)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1522..1541
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1601..1617)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1601..1617)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1614..1636)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1625..1641
                     /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(1649..1655,1656..1673,1674..1679))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1694..1715
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1832..1849)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2003..2591)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2083..2102)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2762..3553,3554..3622))
                     /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     3385..3404
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3623..3727)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3802..3821)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3993..4372
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4373..4576
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4527..4547
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4621..4640
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4621..4639
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4672..4696
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(6800..6824)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(6835..6837,6838..6840,6841..7554)
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(6868..6896)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     7491..7511
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(7576..7593)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7582..7806
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(7852..8229,1..51)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7939..7958)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8149..8170
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
       61 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
      121 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
      181 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
      241 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
      301 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
      361 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
      421 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
      481 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
      541 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
      601 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
      661 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
      721 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
      781 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
      841 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
      901 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
      961 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     1021 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     1081 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     1141 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     1201 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     1261 actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     1321 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     1381 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     1441 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     1501 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     1561 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     1621 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     1681 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     1741 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     1801 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     1861 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     1921 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1981 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2041 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2101 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2161 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2221 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2281 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2341 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2401 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     2461 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2521 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2581 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2641 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2701 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     2761 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     2821 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     2881 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     2941 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     3001 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     3061 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     3121 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     3181 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     3241 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     3301 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     3361 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     3421 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     3481 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     3541 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     3601 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     3661 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     3721 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     3781 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     3841 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     3901 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
     3961 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
     4021 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     4081 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     4141 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     4201 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     4261 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     4321 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     4381 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     4441 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     4501 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     4561 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat
     4621 taatacgact cactataggg agacccaagc tggctagtta agcttgatca aacaagtttg
     4681 tacaaaaaag caggctccac catgctgccc gctcgcctac cgttcaggct gctgagcctt
     4741 ttccttcgtg gatccgctcc cacggcagcg cgccatggcc tccgggagcc gctcctggag
     4801 aggaggtgcg ctgccgcctc ctccttccag cactcatcga gtctgggacg cgagcttcct
     4861 tatgaccccg tggacacgga gggctttgga gaaggtggtg acatgcagga gcgttttctg
     4921 ttcccggagt acatcctgga tccggagccg caacccaccc gcgaaaagca gctgcaggag
     4981 ctccagcaac agcaggagga ggaggagcga cagaggcagc agcggcggga ggagcggcga
     5041 cagcaaaacc tacgggccag gtcccgggag cacccggtcg tggggcaccc ggacccggca
     5101 ttgccgccca gcggcgtgaa ctgctcgggc tgcggggcag agctgcactg ccaggacgcc
     5161 ggagtgcccg gctacctgcc ccgagagaag ttcctccgca cggcggaggc agacggcggg
     5221 ctggcacgga ccgtgtgcca gcgctgctgg ctgctgtcgc accaccggcg cgctctacgc
     5281 ctgcaggtga gccgcgagca gtacctggag ctggtgagcg ccgcgttgcg gcggcccggc
     5341 ccctccctgg tgctctacat ggtggacctg ctggacctgc ccgacgccct gctgcccgac
     5401 ttgcccgcgc tggtgggccc caagcagctg atcgtgctgg gaaacaaagt ggacctcctg
     5461 ccccaggatg ctcctggcta ccggcagagg ctgcgggagc gactgtggga ggactgtgcc
     5521 cgcgccgggc tcctgctggc ccctggccac caagggccac agcgccccgt caaggacgag
     5581 ccacaggacg gggagaatcc gaatccgccg aactggtccc gcacagtggt cagggacgtg
     5641 cggctgatca gcgccaagac cggctatgga gtggaagagt tgatctctgc ccttcagcgc
     5701 tcctggcgct accgtgggga cgtctactta gtgggcgcca ccaacgccgg caaatccact
     5761 ctctttaaca cgctcctgga gtccgattac tgcactgcca agggctccga ggccatcgac
     5821 agagccacca tctccccttg gccaggtact acattaaacc ttctgaagtt tcctatttgc
     5881 aacccaactc cttacagaat gtttaaaagg catcaaagac ttaaaaaaga ttcaactcaa
     5941 gctgaagaag atcttagtga gcaagaacaa aatcagctta atgtcctcaa aaagcatggt
     6001 tatgtcgtag gaagagttgg aaggacattc ttgtattcag aagaacagaa ggataacatt
     6061 ccctttgagt ttgatgctga ttcacttgcc tttgacatgg aaaatgaccc tgttatgggt
     6121 acacacaaat ccaccaaaca agtagaattg actgcacaag atgtgaaaga tgcccactgg
     6181 ttttatgaca cccctggaat tacaaaagaa aattgtattt taaatcttct aacagaaaaa
     6241 gaagtaaata ttgttttgcc aacacagtcc attgttccaa gaacttttgt gcttaaacca
     6301 ggaatggttc tgtttttggg tgctataggc cgcatagatt tcctgcaggg aaatcagtca
     6361 gcttggttta cagtcgtggc ttccaacatc ctccctgtgc atatcacctc cttggacagg
     6421 gcagacgctc tgtatcagaa gcatgcaggt catacgttac tccagattcc aatgggtgga
     6481 aaagaacgaa tggcaggatt tcctcctctt gttgctgaag acattatgtt aaaagaagga
     6541 ctgggggcat ctgaagcagt ggccgacatc aagttttcct ctgcaggttg ggtttcagta
     6601 acacctaatt ttaaggacag actgcatctc cgaggctata cacctgaagg aacagttttg
     6661 accgtccggc cccctctctt gccatatatt gttaacatca aaggacagcg catcaagaaa
     6721 agtgtggcct ataaaaccaa gaagcctcct tcccttatgt acaacgtgag gaagaagaaa
     6781 ggaaagataa atgtattgga cccagctttc ttgtacaaag tggttcgatc tagaatggct
     6841 agcaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat
     6901 gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgctac atacggaaag
     6961 cttaccctta aatttatttg cactactgga aaactacctg ttccatggcc aacacttgtc
     7021 actactttct cttatggtgt tcaatgcttt tcccgttatc cggatcatat gaaacggcat
     7081 gactttttca agagtgccat gcccgaaggt tatgtacagg aacgcactat atctttcaaa
     7141 gatgacggga actacaagac gcgtgctgaa gtcaagtttg aaggtgatac ccttgttaat
     7201 cgtatcgagt taaaaggtat tgattttaaa gaagatggaa acattctcgg acacaaactc
     7261 gagtacaact ataactcaca caatgtatac atcacggcag acaaacaaaa gaatggaatc
     7321 aaagctaact tcaaaattcg ccacaacatt gaagatggat ccgttcaact agcagaccat
     7381 tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa ccattacctg
     7441 tcgacacaat ctgccctttc gaaagatccc aacgaaaagc gtgaccacat ggtccttctt
     7501 gagtttgtaa ctgctgctgg gattacacat ggcatggatg agctctacaa ataatgaatt
     7561 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     7621 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     7681 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     7741 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     7801 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     7861 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     7921 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     7981 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     8041 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     8101 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     8161 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     8221 tgccgattt
//
