LOCUS       sequence-225072-        8485 bp    DNA     circular SYN 03-DEC-2025
DEFINITION  Contains an optimized 96-mer TetO repeat for imaging of desired 
            loci.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8485)
  AUTHORS   Tasan I, Sustackova G, Zhang L, Kim J, Sivaguru M, HamediRad M, Wang
            Y, Genova J, Ma J, Belmont AS, Zhao H
  TITLE     CRISPR/Cas9-mediated knock-in of an optimized TetO repeat for live 
            cell imaging of endogenous loci.
  JOURNAL   Nucleic Acids Res. 2018 Jun 15. pii: 5038283. doi: 
            10.1093/nar/gky501.
  PUBMED    29912475
REFERENCE   2  (bases 1 to 8485)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 3, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. 2018 Jun 15. pii: 5038283. doi: 10.1093/nar/gky501."
COMMENT     Sequence Label: pSP2-96-merTetO-EFS-BLaR
FEATURES             Location/Qualifiers
     source          1..8485
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    21..39
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    50..68
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    79..97
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    124..142
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    153..171
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    182..200
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    227..245
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    256..274
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    285..303
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    330..348
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    359..377
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    388..406
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    433..451
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    462..480
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    491..509
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    536..554
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    565..583
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    594..612
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    639..657
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    669..687
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    698..716
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    743..761
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    772..790
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    801..819
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    846..864
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    875..893
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    904..922
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    948..966
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    977..995
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1006..1024
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1051..1069
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1080..1098
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1109..1127
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1152..1170
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1181..1199
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1210..1228
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1255..1273
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1284..1302
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1313..1331
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1358..1376
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1387..1405
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1416..1434
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1461..1479
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1490..1508
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1519..1537
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1564..1582
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1593..1611
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1622..1640
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1667..1685
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1696..1714
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1725..1743
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1770..1788
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1799..1817
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1828..1846
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1873..1891
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1902..1920
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1931..1949
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1976..1994
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2005..2023
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2034..2052
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2079..2097
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2108..2126
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2137..2155
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2182..2200
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2211..2229
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2240..2258
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2285..2303
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2314..2332
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2343..2361
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2388..2406
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2417..2435
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2446..2464
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2491..2509
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2520..2538
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2549..2567
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2594..2612
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2623..2641
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2652..2670
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2697..2715
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2726..2744
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2755..2773
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2800..2818
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2829..2847
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2858..2876
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2903..2921
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2932..2950
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2961..2979
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3006..3024
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3035..3053
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3064..3082
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3109..3127
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3138..3156
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3167..3185
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3212..3230
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3241..3259
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3270..3288
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3305..3338
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3390..3601
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3609..3618
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3615..4013
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4342..4476
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    4489..4522
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             4569..4760
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        4819..5030
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      5037..5046
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5043..6173
                     /codon_start=1
                     /product="herpes simplex virus thymidine kinase"
                     /label=HSV TK
                     /note="confers sensitivity to nucleoside analogs such as 
                     acyclovir or ganciclovir"
                     /translation="MASYPCHQHASAFDQAARSRGHSNRRTALRPRRQQEATEVRLEQK
                     MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWQVLGASETIANIYTTQ
                     HRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDR
                     HPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQR
                     PGERLDLAMLAAIRRVYGLLANTVRYLQGGGSWREDWGQLSGTAVPPQGAEPQSNAGPR
                     PHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRPMHVFILDYDQSPAGCRDAL
                     LQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     polyA_signal    6228..6349
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6485..6502)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6656..7244)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6736..6755)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7415..8203)
                     /codon_start=1
                     /gene="bla(M)"
                     /product="beta-lactamase lacking the signal sequence"
                     /label=bla(M)
                     /note="allows cytosolic expression of beta-lactamase"
                     /translation="PETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMST
                     FKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMS
                     DNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPAAMATT
                     LRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAA
                     LGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     primer_bind     8038..8057
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8277..8381)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8449..8467
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgccctagg aatgggttag tccctatcag tgatagagag aagggtgttt ccctatcagt
       61 gatagagatg gtggtgattc cctatcagtg atagagaatt gaaatttact aggagaatag
      121 aagtccctat cagtgataga gaatattggt gttccctatc agtgatagag agggaatagt
      181 ttccctatca gtgatagaga tagtttgggg actagggaag agtttgtccc tatcagtgat
      241 agagagtgtg tatagtccct atcagtgata gagaagaagt aatgtcccta tcagtgatag
      301 agaagttaat gtaactagga aaaatataat ccctatcagt gatagagaaa ttttgagatc
      361 cctatcagtg atagagattg ggagtggtcc ctatcagtga tagagatgga agaaggacta
      421 gggaatgtat gatccctatc agtgatagag aagagttagt gtccctatca gtgatagaga
      481 gtagttggat tccctatcag tgatagagat aggggtgata ctaggtgggg gattgtccct
      541 atcagtgata gagaggaaaa aggttcccta tcagtgatag agagttggtg gaatccctat
      601 cagtgataga gagtaaatgt taactaggaa gatgagtttc cctatcagtg atagagattg
      661 tatgaagttc cctatcagtg atagagatta tttaggttcc ctatcagtga tagagaggtg
      721 agttggacta ggtaatagat ggtccctatc agtgatagag aggggtattg gtccctatca
      781 gtgatagaga gtatagttat tccctatcag tgatagagaa gatagtgata ctaggaggag
      841 ttgggtccct atcagtgata gagattagag atggtcccta tcagtgatag agagtaggaa
      901 atttccctat cagtgataga gataaagggt gactaggaag tattgaatcc ctatcagtga
      961 tagagaatat attgtgtccc tatcagtgat agagagtagg gtggttccct atcagtgata
     1021 gagaaggtgg ggatactagg tgggtgatat tccctatcag tgatagagat tatagagtgt
     1081 ccctatcagt gatagagata gggttgattc cctatcagtg atagagaagg tttggaaact
     1141 aggtagaggt atccctatca gtgatagaga ggtgttaggg tccctatcag tgatagagag
     1201 gataaagtgt ccctatcagt gatagagatg ggttaaagac taggtagtgg aaagtcccta
     1261 tcagtgatag agagattttt atttccctat cagtgataga gagaggggat aatccctatc
     1321 agtgatagag attgtagagt tactagggaa gtggttgtcc ctatcagtga tagagagtag
     1381 agggagtccc tatcagtgat agagaaaagg tggtttccct atcagtgata gagaataagt
     1441 tagtactagg gtgtaattta tccctatcag tgatagagag tatagagggt ccctatcagt
     1501 gatagagagt aatatggttc cctatcagtg atagagatgg aggtgggact aggagtttag
     1561 atttccctat cagtgataga gaggtaattt tgtccctatc agtgatagag attagtatgg
     1621 ttccctatca gtgatagaga gggttgtgat actagggagg tggagatccc tatcagtgat
     1681 agagagtaag gtagttccct atcagtgata gagaggtgaa ggtgtcccta tcagtgatag
     1741 agatatatag gaaactaggg gagtagttgt ccctatcagt gatagagagt atggaggttc
     1801 cctatcagtg atagagatgg gaaaaagtcc ctatcagtga tagagagata gtggggacta
     1861 ggaatagatt tatccctatc agtgatagag attttaattt ttccctatca gtgatagaga
     1921 gaggttagat tccctatcag tgatagagag gaggtggaga ctagggttaa gataatccct
     1981 atcagtgata gagatgtatg ggtatcccta tcagtgatag agatgggggg tggtccctat
     2041 cagtgataga gatgaaagtg agactaggaa gttatgagtc cctatcagtg atagagatga
     2101 aaattattcc ctatcagtga tagagatttg agagtatccc tatcagtgat agagaataat
     2161 tgaatactag ggaattgtag ttccctatca gtgatagaga ttatgggggg tccctatcag
     2221 tgatagagat gtggtgtggt ccctatcagt gatagagaaa agattgtaac taggttatgt
     2281 tgtatcccta tcagtgatag agagagaaga taatccctat cagtgataga gagggagatg
     2341 aatccctatc agtgatagag agtaaaagtt tactagggat tttagattcc ctatcagtga
     2401 tagagaagtg gggatgtccc tatcagtgat agagatgtag tatattccct atcagtgata
     2461 gagagtttta ggaaactagg aagtgtatat tccctatcag tgatagagaa gagatgttgt
     2521 ccctatcagt gatagagaaa gggtgggttc cctatcagtg atagagagta atttataact
     2581 aggtgatgtt ggttccctat cagtgataga gaaaggtaga tatccctatc agtgatagag
     2641 atgggagagt ttccctatca gtgatagaga agatgttaag actaggttag tgggtgtccc
     2701 tatcagtgat agagataaag gtgtgtccct atcagtgata gagaggatat atgttcccta
     2761 tcagtgatag agaatggaat tggactaggt gttgtttggt ccctatcagt gatagagagt
     2821 ggttgatttc cctatcagtg atagagagta ttgttagtcc ctatcagtga tagagagtta
     2881 atttggacta ggtaggaata ggtccctatc agtgatagag aatgggtttt atccctatca
     2941 gtgatagaga tagggtaaat tccctatcag tgatagagat taagataata ctaggtagta
     3001 ggtgttccct atcagtgata gagattgtat ttggtcccta tcagtgatag agagggggtg
     3061 tgttccctat cagtgataga gaattgatag atactagggt atatatgttc cctatcagtg
     3121 atagagaaat ttaaaagtcc ctatcagtga tagagaattg gggttgtccc tatcagtgat
     3181 agagatgttt tatatactag ggagatgaat ttccctatca gtgatagaga tgggtttgag
     3241 tccctatcag tgatagagag gaaaaggatt ccctatcagt gatagagata ggttgatgac
     3301 tagtataact tcgtataatg tatgctatac gaagttatca tatggctagg tcttgaaagg
     3361 agtgggaatt ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga
     3421 gaagttgggg ggaggggtcg gcaattgatc cggtgcctag agaaggtggc gcggggtaaa
     3481 ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta
     3541 tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca
     3601 ggaagcttgc caccatggcc aagcctttgt ctcaagaaga atccaccctc attgaaagag
     3661 caacggctac aatcaacagc atccccatct ctgaagacta cagcgtcgcc agcgcagctc
     3721 tctctagcga cggccgcatc ttcactggtg tcaatgtata tcattttact gggggacctt
     3781 gcgcagaact cgtggtgctg ggcactgctg ctgctgcggc agctggcaac ctgacttgta
     3841 tcgtcgcgat cggaaatgag aacaggggca tcttgagccc ctgcggacgg tgccgacagg
     3901 ttcttctcga tctgcatcct gggatcaaag ccatagtgaa ggacagtgat ggacagccga
     3961 cggcagttgg gattcgtgaa ttgctgccct ctggttatgt gtgggagggc taacccttat
     4021 acgcaaggga gtagatgccg accgaacaag agctgatttc gagaacgcct cagccagcaa
     4081 ctcgcgcgag cctagcaagg caaatgcgag agaacggcct tacgcttggt ggcacagttc
     4141 tcgtccacag ttcgctaagc tcgctcggct gggtcgcggg agggccggtc gcagtgattc
     4201 aggcccttct ggattgtgtt ggtccccagg gcacgattgt catgcccacg cactcgggtg
     4261 atctgactga tcccgcagat tggagatcgc cgcccgtgcc tgccgattgg gtgcagatca
     4321 gcctcgaggc cagctagctt gaacttgttt attgcagctt ataatggtta caaataaagc
     4381 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4441 tccaaactca tcaatgtatc ttatcatgtc tggatcacat gtttcgaaat aacttcgtat
     4501 aatgtatgct atacgaagtt atctcgagag cgctgctagc ttaaggtacc atcgatagat
     4561 ctgggcccgt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt atcagaagcc
     4621 agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag gcagacatct
     4681 gtgaatcgct tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg cgtttcggtg
     4741 atgacggtga aaacctctga cttaagcctg agggctaggt cttgaaagga gtgggaattg
     4801 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
     4861 gaggggtcgg caattgatcc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
     4921 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
     4981 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gggcgcgcca
     5041 ccatggcttc gtacccctgc catcaacacg cgtctgcgtt cgaccaggct gcgcgttctc
     5101 gcggccatag caaccgacgt acggcgttgc gccctcgccg gcagcaagaa gccacggaag
     5161 tccgcctgga gcagaaaatg cccacgctac tgcgggttta tatagacggt cctcacggga
     5221 tggggaaaac caccaccacg caactgctgg tggccctggg ttcgcgcgac gatatcgtct
     5281 acgtacccga gccgatgact tactggcagg tgctgggggc ttccgagaca atcgcgaaca
     5341 tctacaccac acaacaccgc ctcgaccagg gtgagatatc ggccggggac gcggcggtgg
     5401 taatgacaag cgcccagata acaatgggca tgccttatgc cgtgaccgac gccgttctgg
     5461 ctcctcatat cgggggggag gctgggagct cacatgcccc gcccccggcc ctcaccctca
     5521 tcttcgaccg ccatcccatc gccgccctcc tgtgctaccc ggccgcgcga taccttatgg
     5581 gcagcatgac cccccaggcc gtgctggcgt tcgtggccct catcccgccg accttgcccg
     5641 gcacaaacat cgtgttgggg gcccttccgg aggacagaca catcgaccgc ctggccaaac
     5701 gccagcgccc cggcgagcgg cttgacctgg ctatgctggc cgcgattcgc cgcgtttacg
     5761 ggctgcttgc caatacggtg cggtatctgc agggcggcgg gtcgtggcgg gaggattggg
     5821 gacagctttc ggggacggcc gtgccgcccc agggtgccga gccccagagc aacgcgggcc
     5881 cacgacccca tatcggggac acgttattta ccctgtttcg ggcccccgag ttgctggccc
     5941 ccaacggcga cctgtacaac gtgtttgcct gggccttgga cgtcttggcc aaacgcctcc
     6001 gtcccatgca cgtctttatc ctggattacg accaatcgcc cgccggctgc cgggacgccc
     6061 tgctgcaact tacctccggg atggtccaga cccacgtcac cacccccggc tccataccga
     6121 cgatctgcga cctggcgcgc acgtttgccc gggagatggg ggaggctaac tgacgatgat
     6181 aacgcgactc tagagtcggg gcggccggcc gcttcgagca gacatgataa gatacattga
     6241 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     6301 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     6361 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     6421 aaacctctac aaatgtggta aaatcgataa ggatccgttt gcgtattggg cgctcttccg
     6481 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     6541 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     6601 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6661 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6721 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6781 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6841 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6901 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6961 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     7021 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     7081 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     7141 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     7201 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     7261 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     7321 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     7381 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     7441 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     7501 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     7561 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     7621 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     7681 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct gcaggcatcg
     7741 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     7801 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     7861 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7921 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     7981 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata
     8041 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     8101 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     8161 ccaactgatc ttcagcatct tttactttca ccagcgtttc tggagtgagc aaaaacagga
     8221 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     8281 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     8341 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     8401 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     8461 acgaggccct ttcgtcttca agaat
//
