LOCUS       sequence-126268-        5009 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  OT-II TCR beta chain cDNA in pCRII.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5009)
  AUTHORS   Barnden MJ, Allison J, Heath WR, Carbone FR
  TITLE     Defective TCR expression in transgenic mice constructed using 
            cDNA-based alpha- and beta-chain genes under the control of 
            heterologous regulatory elements.
  JOURNAL   Immunol Cell Biol. 1998 Feb;76(1):34-40.
  PUBMED    9553774
REFERENCE   2  (bases 1 to 5009)
  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: "Immunol 
            Cell Biol."; date: "1998-02"; volume: "76(1)"; pages: "34-40"
COMMENT     Sequence Label: pCRII-1.1-2.01 beta
FEATURES             Location/Qualifiers
     source          1..5009
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        239..257
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     239..256
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1442..1461)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1443..1461)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1468..1485)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1468..1484)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1477..1499)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1625..2053
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1712..1731)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1922..1943
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             2397..3191
                     /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(2451..2470)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3061..3080
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3209..3277,3278..4069)
                     /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     complement(3427..3446)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4240..4828
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4729..4748
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4982..4999
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat
      241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcca
      301 ctagtaacgg ccgccagtgt gctggaattc ggcttggatc cagcatgtct aacactgcct
      361 tccctgaccc cgcctggaac accaccctgc tatcttgggt tgctctcttt ctcctgggaa
      421 caagttcagc aaattctggg gttgtccagt ctccaagata cataatcaaa ggaaagggag
      481 aaaggtccat tctaaaatgt attcccatct ctggacatct ctctgtggcc tggtatcaac
      541 agactcaggg gcaggaacta aagttcttca ttcagcatta tgataaaatg gagagagata
      601 aaggaaacct gcccagcaga ttctcagtcc aacagtttga tgactatcac tctgagatga
      661 acatgagtgc cttggagcta gaggactctg ccgtgtactt ctgtgccagc tctctcgggg
      721 gggagagtca aaacaccttg tactttggtg cgggcacccg actatcggtg ctagaggatc
      781 tgagaaatgt gactccaccc aaggtctcct tgtttgagcc atcaaaagca gagattgcaa
      841 acaaacaaaa ggctaccctc gtgtgcttgg ccaggggctt cttccctgac cacgtggagc
      901 tgagctggtg ggtgaatggc aaggaggtcc acagtggggt cagcacggac cctcaggcct
      961 acaaggagag caattatagc tactgcctga gcagccgcct gagggtctct gctaccttct
     1021 ggcacaatcc tcgaaaccac ttccgctgcc aagtgcagtt ccatgggctt tcagaggagg
     1081 acaagtggcc agagggctca cccaaacctg tcacacagaa catcagtgca gaggcctggg
     1141 gccgagcaga ctgtggaatc acttcagcat cctatcatca gggggttctg tctgcaacca
     1201 tcctctatga gatcctactg gggaaggcca ccctatatgc tgtgctggtc agtggcctgg
     1261 tgctgatggc catggtcaag aaaaaaaatt cctgagacaa acttttatgc atcctgagcc
     1321 gttcttcacc ctggccatag attttcctgc accttctcta attcctgaga tctaagccga
     1381 attctgcaga tatccatcac actggcggcc gctcgagcat gcatctagag ggcccaattc
     1441 gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc gtgactggga
     1501 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     1561 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     1621 atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     1681 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     1741 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     1801 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     1861 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     1921 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     1981 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2041 tttaacgcga attttaacaa aattcagggc gcaagggctg ctaaaggaag cggaacacgt
     2101 agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac tgggctatct
     2161 ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg cttacatggc
     2221 gatagctaga ctgggcggtt ttatggacag caagcgaacc ggaattgcca gctggggcgc
     2281 cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga
     2341 tctgatggcg caggggatca agatctgatc aagagacagg atgaggatcg tttcgcatga
     2401 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     2461 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     2521 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     2581 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     2641 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     2701 tcctgtcatc ccaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     2761 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     2821 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     2881 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     2941 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3001 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     3061 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     3121 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     3181 agttcttctg aattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     3241 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     3301 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     3361 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     3421 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     3481 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     3541 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     3601 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     3661 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     3721 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     3781 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     3841 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     3901 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     3961 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4021 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4081 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4141 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4201 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4261 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4321 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4381 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4441 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4501 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4561 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4621 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4681 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4741 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4801 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4861 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     4921 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     4981 cagcgagtca gtgagcgagg aagcggaag
//
