LOCUS       sequence-365601-        8562 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-2 vector encoding PTetO7-driven SEAP-p2A-iRFP670, PmPGK1-driven
            tTA and PmPGK1-driven YPet expression 
            (PTetO7-CMVmin-2-SEAP-p2A-iRFP670-pA::PmPGK1-tTA-pA::PmPGK1-YPet-pA)
            ..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8562)
  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 8562)
  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: pVH663
FEATURES             Location/Qualifiers
     source          1..8562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(1..792,793..861))
                     /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     624..643
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(862..966)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    1047..1065
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1083..1101
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1119..1137
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1155..1173
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1191..1209
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1227..1245
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1263..1281
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1335..1359
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1411..1430
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1411..1429
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3060..3116
                     /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"
     CDS             3123..4052
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 670 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP670
                     /note="derived from Rhodopseudomonas palustris RpBphP6"
                     /translation="ARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAGA
                     FFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLHR
                     HDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVML
                     YRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSRI
                     VPEHDASGAALDLSFAHLRSISPCHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYEP
                     RAVPMAQRVAAEMFADFLSLHFTAAHHQR"
     primer_bind     complement(4113..4130)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4119..4343
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        4391..4890
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             join(4943..5560,5561..5944)
                     /codon_start=1
                     /product="tetracycline-controlled transactivator, 
                     comprising a fusion of the tetracycline repressor TetR with
                     the C-terminal activation domain of herpes simplex virus 
                     VP16"
                     /label=tTA
                     /note="In the Tet-Off(R) system, tTA binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="SRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHVK
                     NKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTRP
                     TEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTTD
                     SMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTIE
                     GLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHAD
                     ALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     complement(5013..5032)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(6072..6088)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6072..6088)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6128..6627
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6995..7014)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    7545..7566
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7683..7700)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7854..8442)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7934..7953)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
       61 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
      121 cagtgctgca atgataccgc ggctcccacg ctcaccggct ccagatttat cagcaataaa
      181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
      241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
      301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
      361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
      421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
      481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
      541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
      601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
      661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
      721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
      781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
      841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
      901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
      961 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccaaga tctccacgcg
     1021 tccgtacacg cctaaagcat atacgttctc tatcactgat agggagtaaa ctggatatac
     1081 gttctctatc actgataggg agtaaactgt agatacgttc tctatcactg atagggagta
     1141 aactggtcat acgttctcta tcactgatag ggagtaaact ccttatacgt tctctatcac
     1201 tgatagggag taaagtctgc atacgttctc tatcactgat agggagtaaa ctcttcatac
     1261 gttctctatc actgataggg agtaaactcg agtatgtcga ggtggcgtgt acggtgggag
     1321 gcctatataa gcagagctcg tttagtgaac cgtcagatcg cgagctctct ggctaactag
     1381 agaacccact gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc
     1441 tgttcgaagc ggaattcacc atgactagtt ggctgtggct ggtgctgctg ctgctgtgcc
     1501 tggccggcaa cgtgcaggcc gctagtatca tccccgtgga ggaggagaac cccgacttct
     1561 ggaacagaga ggccgccgag gccctgggcg ccgccaagaa gctgcagccc gcccagaccg
     1621 ccgccaagaa cctgatcatc ttcctgggcg acggcatggg cgtgagcacc gtgaccgccg
     1681 ccagaatcct gaagggccag aagaaggaca agctgggccc cgagatcccc ctggccatgg
     1741 acagattccc ctacgtggcc ctgagcaaga cctacaacgt ggacaagcac gtgcccgaca
     1801 gcggcgccac cgccaccgcc tacctgtgcg gcgtgaaggg caacttccag accatcggcc
     1861 tgagcgccgc cgccagattc aaccagtgca acaccaccag aggcaacgag gtgatcagcg
     1921 tgatgaacag agccaagaag gccggcaaga gcgtgggcgt ggtgaccacc accagagtgc
     1981 agcacgccag ccccgccggc acctacgccc acaccgtgaa cagaaactgg tacagcgacg
     2041 ccgacgtgcc cgccagcgcc agacaggagg gctgccagga catcgccacc cagctgatca
     2101 gcaacatgga catcgacgtg atcctgggcg gcggcagaaa gtacatgttc agaatgggca
     2161 cccccgaccc cgagtacccc gacgactaca gccagggcgg caccagactg gacggcaaga
     2221 acctggtgca ggagtggctg gccaagagac agggcgccag atacgtgtgg aacagaaccg
     2281 agctgatgca ggccagcctg gaccccagcg tgacccacct gatgggcctg ttcgagcccg
     2341 gcgacatgaa gtacgagatc cacagagaca gcaccctgga ccccagcctg atggagatga
     2401 ccgaggccgc cctgagactg ctgagcagaa accccagagg cttcttcctg ttcgtggagg
     2461 gcggcagaat cgaccacggc caccacgaga gcagagccta cagagccctg accgagacca
     2521 tcatgttcga cgacgccatc gagagagccg gccagctgac cagcgaggag gacaccctga
     2581 gcctggtgac cgccgaccac agccacgtgt tcagcttcgg cggctacccc ctgagaggca
     2641 gcagcatctt cggcctggcc cccggcaagg ccagagacag aaaggcctac accgtgctgc
     2701 tgtacggcaa cggccccggc tacgtgctga aggacggcgc cagacccgac gtgaccgaga
     2761 gcgagagcgg cagccccgag tacagacagc agagcgccgt gcccctggac gaggagaccc
     2821 acgccggcga ggacgtggcc gtgttcgcca gaggccccca ggcccacctg gtgcacggcg
     2881 tgcaggagca gaccttcatc gcccacgtga tggccttcgc cgcctgcctg gagccctaca
     2941 ccgcctgcga cctggccccc cccgccggca ccaccgacgc cgcccacccc ggctacagca
     3001 gagtgggcgc cgccggcaga ttcgagcaga ccgctagtgg tggttctggt ggaagcggag
     3061 ctactaactt cagcctgctg aagcaggctg gtgacgtgga ggagaaccct ggacctgcta
     3121 gtgcgcgtaa ggtcgatctc acctcctgcg atcgcgagcc gatccacatc cccggcagca
     3181 ttcagccgtg cggctgcctg cttgcctgcg acgcgcaggc ggtgcggatc acgcgcatta
     3241 cggaaaatgc cggcgcgttc tttggacgcg aaactccgcg ggtcggtgag ctactcgccg
     3301 attacttcgg cgagaccgaa gcccatgcgc tgcgcaacgc actggcgcag tcctccgatc
     3361 caaagcgacc ggcgctgatc ttcggttggc gcgacggcct gaccggccgc accttcgaca
     3421 tctcactgca tcgccatgac ggtacatcga tcatcgagtt cgagcctgcg gcggccgaac
     3481 aggccgacaa tccgctgcgg ctgacgcggc agatcatcgc gcgcaccaaa gaactgaagt
     3541 cgctcgaaga gatggccgca cgggtgccgc gctatctgca ggcgatgctc ggctatcacc
     3601 gcgtgatgtt gtaccgcttc gcggacgacg gctccgggat ggtgatcggc gaggcgaagc
     3661 gcagcgacct cgagagcttt ctcggtcagc actttccggc gtcgctggtc ccgcagcagg
     3721 cgcggctact gtacttgaag aacgcgatcc gcgtggtctc ggattcgcgc ggcatcagca
     3781 gccggatcgt gcccgagcac gacgcctccg gcgccgcgct cgatctgtcg ttcgcgcacc
     3841 tgcgcagcat ctcgccctgc catctcgaat ttctgcggaa catgggcgtc agcgcctcga
     3901 tgtcgctgtc gatcatcatt gacggcacgc tatggggatt gatcatctgt catcattacg
     3961 agccgcgtgc cgtgccgatg gcgcagcgcg tcgcggccga aatgttcgcc gacttcttat
     4021 cgctgcactt caccgccgcc caccaccaac gcgctagcgg atccaccggt gtctagaaag
     4081 ctttgaggcc ggcctgcagg cccgctgatc agcctcgact gtgccttcta gttgccagcc
     4141 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     4201 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     4261 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     4321 tggggatgcg gtgggctcta tggtagctta ggaggcggaa agaaccagcg cgtggtaccc
     4381 tcgagctacc gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca
     4441 gccccgctgg gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc
     4501 accggtaggc gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct
     4561 cccctagtca ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg
     4621 gaagtagcac gtctcagtag tctcgtgcag atggacagca ccgctgagca atggaagcgg
     4681 gtaggccttt ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg
     4741 ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc
     4801 cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg
     4861 ttctcctctt cctcatctcc gggcctttcg acctccagcc cgagctcggt ataggttcga
     4921 agcggaattc accatgacta gttccagatt agataaaagt aaagtgatta acagcgcatt
     4981 agagctgctt aatgaggtcg gaatcgaagg tttaacaacc cgtaaactcg cccagaagct
     5041 aggtgtagag cagcctacat tgtattggca tgtaaaaaat aagcgggctt tgctcgacgc
     5101 cttagccatt gagatgttag ataggcacca tactcacttt tgccctttag aaggggaaag
     5161 ctggcaagat tttttacgta ataacgctaa aagttttaga tgtgctttac taagtcatcg
     5221 cgatggagca aaagtacatt taggtacacg gcctacagaa aaacagtatg aaactctcga
     5281 aaatcaatta gcctttttat gccaacaagg tttttcacta gagaatgcat tatatgcact
     5341 cagcgctgtg gggcatttta ctttaggttg cgtattggaa gatcaagagc atcaagtcgc
     5401 taaagaagaa agggaaacac ctactactga tagtatgccg ccattattac gacaagctat
     5461 cgaattattt gatcaccaag gtgcagagcc agccttctta ttcggccttg aattgatcat
     5521 atgcggatta gaaaaacaac ttaaatgtga aagtgggtcc gcgtacagcc gcgcgcgtac
     5581 gaaaaacaat tacgggtcta ccatcgaggg cctgctcgat ctcccggacg acgacgcccc
     5641 cgaagaggcg gggctggcgg ctccgcgcct gtcctttctc cccgcgggac acacgcgcag
     5701 actgtcgacg gcccccccga ccgatgtcag cctgggggac gagctccact tagacggcga
     5761 ggacgtggcg atggcgcatg ccgacgcgct agacgatttc gatctggaca tgttggggga
     5821 cggggattcc ccgggtccgg gatttacccc ccacgactcc gccccctacg gcgctctgga
     5881 tatggccgac ttcgagtttg agcagatgtt taccgatgcc cttggaattg acgagtacgg
     5941 tggggctagt gctagcggat ccaccggtgt ctagaaagct atacaggtgc accgctttat
     6001 aataaaactg atacagagaa cgttatcatg tgtgttggtt ttttttgtgt gtgcggttta
     6061 agggagcctc ggtcatagct gtttcctgtg taaaacgacg gccagcgcgt ggtaccctcg
     6121 agctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc
     6181 ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc
     6241 ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc
     6301 ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa
     6361 gtagcacgtc tcagtagtct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta
     6421 ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg
     6481 ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc gggcgcccga
     6541 aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc
     6601 tcctcttcct catctccggg cctttcgacc tccagcccga gctcggtata ggttcgaagc
     6661 ggaattcacc atgactagtg gtggttctgg tgtgagcaaa ggcgaagagc tgttcaccgg
     6721 cgtggtgccc atcctggtgg agctggacgg cgacgtgaac ggccacaagt tcagcgtgag
     6781 cggcgagggc gagggcgacg ccacctacgg caagctgacc ctgaagctgc tgtgcaccac
     6841 cggcaagctg cccgtgccct ggcccaccct ggtgaccacc ctgggctacg gcgtgcagtg
     6901 cttcgcccgg taccccgacc acatgaagca gcacgacttc ttcaagagcg ccatgcccga
     6961 gggctacgtg caggagcgga ccatcttctt caaggacgac ggcaactaca agacccgggc
     7021 cgaggtgaag ttcgagggcg acaccctggt gaaccggatc gagctgaagg gcatcgactt
     7081 caaggaggac ggcaacatcc tgggccacaa gctggagtac aactacaaca gccacaacgt
     7141 gtacatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccggcacaa
     7201 catcgaggac ggcggcgtgc agctggccga ccactaccag cagaacaccc ccatcggcga
     7261 cggccccgtg ctgctgcccg acaaccacta cctgagctac cagagcgccc tgttcaagga
     7321 ccccaacgag aagcgggacc acatggtgct gctggagttc ctgaccgccg ccggcatcac
     7381 cgagggcatg gacgagctgt acaaggctag cggatccacc ggtgtctaga aagctcagaa
     7441 attatgagag cgtttaataa agtattgact atggttgaca aaacgtgtgt tggagttttt
     7501 tgtgtgcttt gtcatcaact gagccagtag gtcgagggtg cctaatgagt gagctaactc
     7561 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     7621 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     7681 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     7741 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     7801 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     7861 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     7921 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     7981 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     8041 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     8101 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     8161 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     8221 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     8281 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     8341 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     8401 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     8461 tctacgggga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     8521 taaatcaatc taaagtatat atgagtaaac ttggtctgac ag
//
