LOCUS       sequence-54395-e        4290 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 4290)
  AUTHORS   Kuhlman TE, Cox EC
  TITLE     Site-specific chromosomal integration of large synthetic constructs.
  JOURNAL   Nucleic Acids Res. 2010 Apr;38(6):e92. Epub 2010 Jan 4.
  PUBMED    20047970
REFERENCE   2  (bases 1 to 4290)
  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: "Nucleic 
            Acids Res."; date: "2010-04"; volume: "38(6)"; pages: "e92. Epub 
            2010 Jan 4"
COMMENT     Sequence Label: pTKIP-neo
FEATURES             Location/Qualifiers
     source          1..4290
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             238..429
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(423..445)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    531..671
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     586..605
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(686..703)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(857..1445)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(937..956)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1616..2407,2408..2476))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2239..2258
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2477..2581)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2667..2683
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2668..2684
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2706..2739
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     promoter        2803..3302
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2838..2861)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3235..3254
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     promoter        3314..3361
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3380..4183
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3443..3462)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4053..4072
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     protein_bind    4214..4247
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
ORIGIN
        1 gcaggatgct gctggctacc ctgtggaaca cctacatctg tattaacgaa gcgctggcat
       61 tgaccctgag tgatttttct ctggtcccgc cgcatccata ccgccagttg tttaccctca
      121 caacgttcca gtaaccgggc atgttcatca tcagtaaccc gtatcgtgag catcctctct
      181 cgtttcatcg gtatcattac ccccatgaac agaaatcccc cttacacgga ggcatcagtg
      241 accaaacagg aaaaaaccgc ccttaacatg gcccgcttta tcagaagcca gacattaacg
      301 cttctggaga aactcaacga gctggacgcg gatgaacagg cagacatctg tgaatcgctt
      361 cacgaccacg ctgatgagct ttaccgcagc tgcctcgcgc gtttcggtga tgacggtgaa
      421 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
      481 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg
      541 acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga
      601 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
      661 accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
      721 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
      781 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
      841 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
      901 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
      961 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1021 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1081 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1141 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1201 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     1261 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     1321 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1381 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     1441 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     1501 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     1561 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     1621 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     1681 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     1741 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     1801 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     1861 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     1921 ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     1981 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     2041 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     2101 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     2161 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     2221 gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     2281 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     2341 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     2401 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     2461 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     2521 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     2581 gcacatttcc ccgaaaagtg ccaccttagg gataacaggg taattacggc cccaaggtcc
     2641 aaacggtgaa ggtaccgggc cccccctcga ggtcgacggt atcgataagc ttgatatcga
     2701 attccgaagt tcctattctc tagaaagtat aggaacttca ggtctgaaga ggagtttacg
     2761 tccagccaag ctagcttggc tgcaggtcgt cgaaattcta ccgggtaggg gaggcgcttt
     2821 tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa
     2881 gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct
     2941 ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc
     3001 cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc
     3061 agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc
     3121 agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc
     3181 tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct
     3241 gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt
     3301 cgacctgcag cctgttgaca attaatcatc ggcatagtat atcggcatag tataatacga
     3361 caaggtgagg aactaaacca tgggatcggc cattgaacaa gatggattgc acgcaggttc
     3421 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3481 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3541 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     3601 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3661 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     3721 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     3781 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     3841 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     3901 cgccaggctc aaggcgcgca tgcccgacgg cgatgatctc gtcgtgaccc atggcgatgc
     3961 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4021 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4081 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4141 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgaggggatc aattctctag
     4201 agctcgctga tcagaagttc ctattctcta gaaagtatag gaacttcgat ggcgcctcat
     4261 ccctgaagcc aatagggata acagggtaat
//
