LOCUS       sequence-446216-        7810 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses full length GalNAcT2-msGFP2 mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7810)
  TITLE     Yellow and oxidation-resistant derivatives of a monomeric 
            superfolder GFP
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7810)
  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            "Unpublished"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: pEF-GalNAcT2-msGFP2
FEATURES             Location/Qualifiers
     source          1..7810
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      96..231
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     158..177
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             271..1065
                     /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="MVEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(325..344)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     935..954
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1239..1360
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1276..1295)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1330..1349
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1393..1410)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1564..2152)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1644..1663)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2323..3114,3115..3183))
                     /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     2946..2965
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3184..3288)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3364..4526
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3595..4517
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4474..4494
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             6277..6984
                     /codon_start=1
                     /product="photostable monomeric superfolder GFP (Valbuena 
                     et al., 2020)"
                     /label=msGFP2
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTITFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITGGSGGS"
     primer_bind     complement(6319..6340)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     6927..6948
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7008..7025)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7014..7238
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7284..7712
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7371..7390)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7581..7602
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7721..7741)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(7726..7810,1..245)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
       61 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
      121 ccgcccattc tccgccccta ggctgactaa ttttttttat ttatgcagag gccgaggccg
      181 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
      241 gcaaaaagct ccccccccgg gaggtccaca atggttgaac aagatggatt gcacgcaggt
      301 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
      361 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
      421 accgacctgt ccggtgccct gaatgaactc caggacgagg cagcgcggct atcgtggctg
      481 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
      541 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
      601 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
      661 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
      721 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
      781 ttcgccaggc tcaaggcgcg tatgcccgac ggcgaggatc tcgtcgtgac tcatggcgat
      841 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
      901 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
      961 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     1021 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     1081 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     1141 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     1201 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     1261 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     1321 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     1381 tctttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1441 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1501 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1561 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1621 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1681 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1741 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1801 ctccaagctg ggctgtatgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     1861 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1921 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     1981 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2041 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2101 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2161 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2221 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2281 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2341 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2401 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2461 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2521 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2581 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     2641 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     2701 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     2761 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     2821 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     2881 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     2941 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3001 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3061 tcgggcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3121 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3181 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3241 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     3301 aaaagtgcca cctgacgtca gatcgacgga tcgggagatc ggtaccgaat tcaagcttcg
     3361 tgaggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg
     3421 gggggagggg tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa
     3481 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt
     3541 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggtaagt
     3601 gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg cgtgccttga
     3661 attacttcca cctggctcca gtacgtgatt cttgatcccg agctggagcc aggggcgggc
     3721 cttgcgcttt aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg
     3781 gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc
     3841 tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg
     3901 taaatgcggg ccaggatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg
     3961 gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga
     4021 gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc
     4081 cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg
     4141 aaagatggcc gcttcccggc cctgctccag ggggctcaaa atggaggacg cggcgctcgg
     4201 gagagcgggc gggtgagtca cccacacaaa ggaaaggggc ctttccgtcc tcagccgtcg
     4261 cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctggagct
     4321 tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca
     4381 ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat
     4441 ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt
     4501 tttttcttcc atttcaggtg tcgtgaacac gtggccaccg ccaccatgag gcggcgctcg
     4561 aggatgctgc tctgcttcgc cttcctgtgg gtgctgggaa tcgcctacta catgtactcg
     4621 ggaggaggat ctgcgctggc cggaggagcg ggaggaggag ccggcaggaa ggaggactgg
     4681 aatgaaattg accccattaa aaagaaagac cttcatcaca gcaatggaga agagaaagca
     4741 caaagcatgg agaccctccc tccagggaaa gtacggtggc cagactttaa ccaggaagct
     4801 tatgttggag ggacgatggt ccgctccggg caggaccctt acgcccgcaa caagttcaac
     4861 caggtggaga gtgataagct tcgaatggat agagccatcc ctgacacccg gcatgaccag
     4921 tgtcagcgga agcagtggcg ggtggatctg ccggccacca gcgtggtgat cacgtttcac
     4981 aatgaagcca ggtcggccct actcaggacc gtggtcagcg tgcttaagaa aagcccgccc
     5041 catctcataa aagaaatcat cttggtggat gactacagca atgatcctga ggacggggct
     5101 ctcttgggga aaattgagaa agtgcgagtt cttagaaatg atcgacgaga aggcctcatg
     5161 cgctcacggg ttcggggggc cgatgctgcc caagccaagg tcctgacctt cctggacagt
     5221 cactgcgagt gtaatgagca ctggctggag cccctcctgg aaagggtggc ggaggacagg
     5281 actcgggttg tgtcacccat catcgatgtc attaatatgg acaactttca gtatgtgggg
     5341 gcatctgctg acttgaaggg cggttttgat tggaacttgg tattcaagtg ggattacatg
     5401 acgcctgagc agagaaggtc ccggcagggg aacccagtcg cccctataaa aacccccatg
     5461 attgctggtg ggctgtttgt gatggataag ttctattttg aagaactggg gaagtacgac
     5521 atgatgatgg atgtgtgggg aggagagaac ctagagatct cgttccgcgt gtggcagtgt
     5581 ggtggcagcc tggagatcat cccgtgcagc cgtgtgggac acgtgttccg gaagcagcac
     5641 ccctacacgt tcccgggtgg cagtggcact gtctttgccc gaaacacccg ccgggcagca
     5701 gaggtctgga tggatgaata caaaaatttc tattatgcag cagtgccttc tgctagaaac
     5761 gttccttatg gaaatattca gagcagattg gagcttagga agaaactcag ctgcaagcct
     5821 ttcaaatggt accttgaaaa tgtctatcca gagttaaggg ttccagacca tcaggatata
     5881 gcttttgggg ccttgcagca gggaactaac tgcctcgaca ctttgggaca ctttgctgat
     5941 ggtgtggttg gagtttatga atgtcacaat gctgggggaa accaggaatg ggccttgacg
     6001 aaggagaagt cggtgaagca catggatttg tgccttactg tggtggaccg ggcaccgggc
     6061 tctcttataa agctgcaggg ctgccgagaa aatgacagca gacagaaatg ggaacagatc
     6121 gagggcaact ccaagctgag gcacgtgggc agcaacctgt gcctggacag tcgcacggcc
     6181 aagagcgggg gcctaagcgt ggaggtgtgt ggcccggccc tttcgcagca gtggaagttc
     6241 acgctcaacc tgcagcaggg aggaagcgga ggaagcatgg atagcactga gagcctgttc
     6301 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     6361 gtgcgcggcg agggcgaggg cgatgccacc aacggcaagc tgaccctgaa gttcatctgc
     6421 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     6481 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     6541 cccgaaggct acgtccagga gcgcaccatc accttcaagg acgacggcac ctacaagacc
     6601 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     6661 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     6721 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc
     6781 cacaacgtgg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     6841 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc
     6901 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     6961 atcactggag gcagtggcgg ttcttagtct agaagctcgc tgatcagcct cgactgtgcc
     7021 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     7081 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     7141 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     7201 caatagcagg catgctgggg atggcccggg ctctatggct tctgaggcgg aaagaaccag
     7261 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     7321 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     7381 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     7441 gctcccttta gggttccgat ttagtgcttt acggcacctc tcccccaaaa aacttgatta
     7501 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     7561 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     7621 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     7681 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     7741 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     7801 tcagcaacca
//
