LOCUS       sequence-364996-        7216 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-1 vector encoding PSV40-driven Citrine-p2A-SEAP expression 
            (PSV40-Citrine-p2A-SEAP-pA). .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7216)
  AUTHORS   Haellman V, Strittmatter T, Bertschi A, Stucheli P, Fussenegger M
  TITLE     A versatile plasmid architecture for mammalian synthetic biology 
            (VAMSyB).
  JOURNAL   Metab Eng. 2021 Jul;66:41-50. doi: 10.1016/j.ymben.2021.04.003. Epub
            2021 Apr 18.
  PUBMED    33857582
REFERENCE   2  (bases 1 to 7216)
  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: "Metab 
            Eng."; date: "2021-07"; volume: "66:41-50. doi"; pages: " 
            10.1016/j.ymben.2021.04.003. Epub 2021 Apr 18"
COMMENT     Sequence Label: pVH64-Tier1-PSV40-Citrine_p2a_SEAP
FEATURES             Location/Qualifiers
     source          1..7216
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             98..892
                     /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(152..171)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     762..781
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1066..1187
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1103..1122)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1157..1176
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1236..1252)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1236..1252)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1249..1271)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1260..1276
                     /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(1284..1290,1291..1308,1309..1314))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1329..1350
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1467..1484)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1638..2226)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1718..1737)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2397..3188,3189..3257))
                     /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     3020..3039
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3258..3362)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3437..3633
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3484..3619
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3684..3703
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3684..3702
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4058..4077)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             4490..4546
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(6200..6217)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6206..6430
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6476..6904
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6563..6582)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6773..6794
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6913..6933)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6918..7216,1..31)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(7098..7216,1..17)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
       61 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
      121 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
      181 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
      241 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
      301 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
      361 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
      421 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
      481 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
      541 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
      601 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
      661 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
      721 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
      781 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
      841 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
      901 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
      961 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     1021 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     1081 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     1141 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     1201 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     1261 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     1321 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     1381 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     1441 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     1501 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     1561 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     1621 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     1681 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     1741 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     1801 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     1861 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     1921 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     1981 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2041 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2101 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2161 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2221 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2281 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     2341 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     2401 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     2461 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     2521 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     2581 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     2641 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     2701 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     2761 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     2821 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     2881 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     2941 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3001 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     3061 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     3121 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     3181 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     3241 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     3301 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     3361 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct ccacgcgtgg
     3421 taccctcgag gcgatctgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3481 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatcg ctgactaatt
     3541 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3601 ggaggctttt ttggaggcct aggcttttgc aaaagagctc tctggctaac tagagaaccc
     3661 actgcttact ggcttatcga aattaatacg actcactata gggagaccca agctgttcga
     3721 agcggaattc accatgacta gtggtggttc tggtgtgagc aagggcgagg agctgttcac
     3781 cggcgtggtg cccatcctgg tggagctgga cggcgacgtg aacggccaca agttcagcgt
     3841 gagcggcgag ggcgagggcg acgccaccta cggcaagctg accctgaagt tcatctgcac
     3901 caccggcaag ctgcccgtgc cctggcccac cctggtgacc accttcggct acggcctgat
     3961 gtgcttcgcc agataccccg accacatgaa gcagcacgac ttcttcaaga gcgccatgcc
     4021 cgagggctac gtgcaggaga gaaccatctt cttcaaggac gacggcaact acaagaccag
     4081 agccgaggtg aagttcgagg gcgacaccct ggtgaacaga atcgagctga agggcatcga
     4141 cttcaaggag gacggcaaca tcctgggcca caagctggag tacaactaca acagccacaa
     4201 cgtgtacatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatcagaca
     4261 caacatcgag gacggcagcg tgcagctggc cgaccactac cagcagaaca cccccatcgg
     4321 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagagcg ccctgagcaa
     4381 ggaccccaac gagaagagag accacatggt gctgctggag ttcgtgaccg ccgccggcat
     4441 caccctgggc atggacgagc tgtacaagac cggtcacatg ggcagcggcg ccaccaactt
     4501 cagcctgctg aagcaggccg gcgacgtgga ggagaacccc ggccccgacg tgcccccctg
     4561 ccagagccag agaagcagag actctagtat gctgctgctg ctgctgctgc tgggcctgag
     4621 actgcagctg agcctgggca tcatccccgt ggaggaggag aaccccgact tctggaacag
     4681 agaggccgcc gaggccctgg gcgccgccaa gaagctgcag cccgcccaga ccgccgccaa
     4741 gaacctgatc atcttcctgg gcgacggcat gggcgtgagc accgtgaccg ccgccagaat
     4801 cctgaagggc cagaagaagg acaagctggg ccccgagatc cccctggcca tggacagatt
     4861 cccctacgtg gccctgagca agacctacaa cgtggacaag cacgtgcccg acagcggcgc
     4921 caccgccacc gcctacctgt gcggcgtgaa gggcaacttc cagaccatcg gcctgagcgc
     4981 cgccgccaga ttcaaccagt gcaacaccac cagaggcaac gaggtgatca gcgtgatgaa
     5041 cagagccaag aaggccggca agagcgtggg cgtggtgacc accaccagag tgcagcacgc
     5101 cagccccgcc ggcacctacg cccacaccgt gaacagaaac tggtacagcg acgccgacgt
     5161 gcccgccagc gccagacagg agggctgcca ggacatcgcc acccagctga tcagcaacat
     5221 ggacatcgac gtgatcctgg gcggcggcag aaagtacatg ttcagaatgg gcacccccga
     5281 ccccgagtac cccgacgact acagccaggg cggcaccaga ctggacggca agaacctggt
     5341 gcaggagtgg ctggccaaga gacagggcgc cagatacgtg tggaacagaa ccgagctgat
     5401 gcaggccagc ctggacccca gcgtgaccca cctgatgggc ctgttcgagc ccggcgacat
     5461 gaagtacgag atccacagag acagcaccct ggaccccagc ctgatggaga tgaccgaggc
     5521 cgccctgaga ctgctgagca gaaaccccag aggcttcttc ctgttcgtgg agggcggcag
     5581 aatcgaccac ggccaccacg agagcagagc ctacagagcc ctgaccgaga ccatcatgtt
     5641 cgacgacgcc atcgagagag ccggccagct gaccagcgag gaggacaccc tgagcctggt
     5701 gaccgccgac cacagccacg tgttcagctt cggcggctac cccctgagag gcagcagcat
     5761 cttcggcctg gcccccggca aggccagaga cagaaaggcc tacaccgtgc tgctgtacgg
     5821 caacggcccc ggctacgtgc tgaaggacgg cgccagaccc gacgtgaccg agagcgagag
     5881 cggcagcccc gagtacagac agcagagcgc cgtgcccctg gacgaggaga cccacgccgg
     5941 cgaggacgtg gccgtgttcg ccagaggccc ccaggcccac ctggtgcacg gcgtgcagga
     6001 gcagaccttc atcgcccacg tgatggcctt cgccgcctgc ctggagccct acaccgcctg
     6061 cgacctggcc ccccccgccg gcaccaccga cgccgcccac cccggctaca gcagagtggg
     6121 cgccgccggc agattcgagc agaccgctag cggatccacc ggtgtctaga aagctttgag
     6181 gccggcctgc agggatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     6241 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     6301 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     6361 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     6421 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     6481 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     6541 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     6601 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     6661 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     6721 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     6781 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     6841 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     6901 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     6961 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     7021 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     7081 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     7141 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     7201 tccagaagta gtgagg
//
