LOCUS       sequence-364608-        6000 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-1 vector encoding PhCMV-driven yellow fluorescent protein YPet 
            fused to a ""self-cleaving"" peptide p2a (PhCMV-p2a_YPet-pA)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6000)
  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 6000)
  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: pTS2388-Tier1-PhCMV-p2a_Ypet
FEATURES             Location/Qualifiers
     source          1..6000
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(58..78)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        63..392
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      243..378
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     305..324
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             459..1253
                     /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(513..532)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1123..1142
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1427..1548
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1464..1483)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1518..1537
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1597..1613)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1597..1613)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1610..1632)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1621..1637
                     /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(1645..1651,1652..1669,1670..1675))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1690..1711
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1828..1845)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1999..2587)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2079..2098)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2758..3549,3550..3618))
                     /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     3381..3400
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3619..3723)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3794..4173
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4174..4377
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4328..4348
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4422..4441
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4422..4440
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4502..4558
                     /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(4880..4899)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(5345..5362)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5351..5575
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(5621..6000,1..49)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5708..5727)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5918..5939
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
       61 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
      121 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
      181 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
      241 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
      301 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
      361 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
      421 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
      481 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
      541 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
      601 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
      661 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
      721 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
      781 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
      841 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
      901 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
      961 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     1021 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     1081 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     1141 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     1201 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     1261 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     1321 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     1381 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     1441 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     1501 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     1561 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     1621 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     1681 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     1741 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     1801 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1861 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     1921 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1981 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2041 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2101 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2161 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2221 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2281 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2341 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2401 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2461 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2521 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2581 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2641 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2701 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2761 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2821 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     2881 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     2941 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3001 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3061 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3121 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3181 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3241 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3301 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     3361 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     3421 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     3481 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     3541 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     3601 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     3661 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     3721 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tccacgcgtg
     3781 gtaccctcga gttgacattg attattgagt agttattaat agtaatcaat tacggggtca
     3841 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     3901 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     3961 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     4021 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     4081 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     4141 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
     4201 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     4261 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     4321 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc
     4381 tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg
     4441 gagacccaag ctgttcgaag cggaattcac catgactagt ggtggttctg gtggaagcgg
     4501 agctactaac ttcagcctgc tgaagcaggc tggtgacgtg gaggagaacc ctggacctgc
     4561 tagtggtggt tctggtgtga gcaaaggcga agagctgttc accggcgtgg tgcccatcct
     4621 ggtggagctg gacggcgacg tgaacggcca caagttcagc gtgagcggcg agggcgaggg
     4681 cgacgccacc tacggcaagc tgaccctgaa gctgctgtgc accaccggca agctgcccgt
     4741 gccctggccc accctggtga ccaccctggg ctacggcgtg cagtgcttcg cccggtaccc
     4801 cgaccacatg aagcagcacg acttcttcaa gagcgccatg cccgagggct acgtgcagga
     4861 gcggaccatc ttcttcaagg acgacggcaa ctacaagacc cgggccgagg tgaagttcga
     4921 gggcgacacc ctggtgaacc ggatcgagct gaagggcatc gacttcaagg aggacggcaa
     4981 catcctgggc cacaagctgg agtacaacta caacagccac aacgtgtaca tcaccgccga
     5041 caagcagaag aacggcatca aggccaactt caagatccgg cacaacatcg aggacggcgg
     5101 cgtgcagctg gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     5161 gcccgacaac cactacctga gctaccagag cgccctgttc aaggacccca acgagaagcg
     5221 ggaccacatg gtgctgctgg agttcctgac cgccgccggc atcaccgagg gcatggacga
     5281 gctgtacaag gctagcggat ccaccggtgt ctagaaagct ttgaggccgg cctgcaggga
     5341 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     5401 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     5461 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     5521 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     5581 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     5641 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     5701 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     5761 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     5821 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     5881 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     5941 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
//
