LOCUS       sequence-44419-e        5706 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 5706)
  AUTHORS   Cortesio CL, Chan KT, Perrin BJ, Burton NO, Zhang S, Zhang ZY, 
            Huttenlocher A
  TITLE     Calpain 2 and PTP1B function in a novel pathway with Src to regulate
            invadopodia dynamics and breast cancer cell invasion.
  JOURNAL   J Cell Biol. 2008 Mar 10;180(5):957-71.
  PUBMED    18332219
REFERENCE   2  (bases 1 to 5706)
  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: "J Cell 
            Biol."; date: "2008-03-10"; volume: "180(5)"; pages: "957-71"
COMMENT     Sequence Label: pTrcHisA-rat-PTP1B
FEATURES             Location/Qualifiers
     source          1..5706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(193..198,199..215,216..222)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    230..246
                     /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     235..257
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    263..332
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region 
                     (Berg et al., 1989)"
     CDS             383..409
                     /codon_start=1
                     /product="synthetic cistron containing a ribosome binding 
                     site (Shine-Dalgarno sequence), for enhancing the bacterial
                     expression of a downstream cistron (Schoner, 1997)"
                     /label=minicistron
                     /note="This first cistron should terminate 3 bp upstream of
                     the ATG for the second cistron."
                     /translation="MYRLNKEE"
     CDS             425..442
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             446..478
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             491..505
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     primer_bind     complement(1889..1906)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2059..2145
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2237..2264
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2284..2375
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2376..2444,2445..3236)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2594..2613)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3407..3995
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3896..3915
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4149..4166
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4181..4321
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4247..4266)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4407..4429
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4507..4584
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             4585..5667
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4604..4623)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
       61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
      121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
      181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagcgccgct gagaaaaagc gaagcggcac tgctctttaa
      301 caatttatca gacaatctgt gtgggcactc gaccggaatt atcgattaac tttattatta
      361 aaaattaaag aggtatatat taatgtatcg attaaataag gaggaataaa ccatgggggg
      421 ttctcatcat catcatcatc atggtatggc tagcatgact ggtggacagc aaatgggtcg
      481 gactctgtac gacgatgacg ataaggatcg atggggatcc atggaaatgg agaaggaatt
      541 cgagcagatc gataaggctg ggaactgggc ggctatttac caggatattc gacatgaagc
      601 cagtgacttc ccatgcagaa tagcgaaact tcctaagaac aaaaaccgga acaggtaccg
      661 agatgtcagc ccttttgacc acagtcggat taaattgcat caggaagata atgactatat
      721 caatgccagc ttgataaaaa tggaggaagc ccagaggagc tatatcctca cccagggccc
      781 tttaccaaac acgtgcgggc acttctggga gatggtgtgg gagcagaaga gcaggggcgt
      841 ggtcatgctc aaccgcatca tggagaaagg ctcgttaaaa tgtgcccagt attggccaca
      901 gaaagaagaa aaagagatgg tcttcgatga caccaatttg aagctgacac tgatctctga
      961 agatgtcaag tcatattaca cagtacggca gttggagttg gagaacctgg ctacccagga
     1021 ggctcgagag atcctgcatt tccactacac cacctggcct gactttggag tccctgagtc
     1081 acctgcctct ttcctcaatt tcctattcaa agtccgagag tcaggctcac tcagcccaga
     1141 gcacggcccc attgtggtcc actgcagtgc tggcattggc aggtcaggga ccttctgcct
     1201 ggctgacacc tgcctcttac tgatggacaa gaggaaagac ccgtcctctg tggacatcaa
     1261 gaaagtgctg ttggagatgc gcaggttccg catggggctc atccagacgg ccgaccaact
     1321 gcgcttctcc tacctggctg tgatcgaggg tgcaaagttc atcatgggcg actcgtcagt
     1381 gcaggatcag tggaaggagc tttcccatga agacctggag cctccccctg agcacgtgcc
     1441 cccacctccc cggccaccca aacgcacatt ggagcctcac aatggcaagt gcaaggagct
     1501 cttctccaac caccagtggg tgagcgagga gagctgtgag gatgaggaca tcctggccag
     1561 agaggaaagc agagccccct caattgctgt gcacagcatg agcagtatga gtcaagacac
     1621 tgaagttagg aaacggatgg tgggtggagg tcttcaaagt gctcaggcat ctgtccccac
     1681 tgaggaagag ctgtccccaa ccgaggagga acaaaaggca cacaggccag ttcactggaa
     1741 gcccttcctg gtcaacgtgt gcatggccac ggccctggcg actggcgcgt acctctgtta
     1801 ccgggtatgt tttcactgaa gatctgcagc tggtaccata tgggaattcg aagcttggct
     1861 gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg
     1921 gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc
     1981 cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag
     2041 tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     2101 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat
     2161 ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc
     2221 aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact
     2281 ctttttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2341 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     2401 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     2461 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     2521 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     2581 cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac
     2641 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     2701 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     2761 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     2821 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     2881 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     2941 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     3001 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     3061 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3121 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     3181 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     3241 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     3301 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     3361 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3421 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3481 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3541 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3601 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3661 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3721 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3781 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3841 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3901 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3961 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4021 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     4081 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     4141 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     4201 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg
     4261 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     4321 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     4381 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     4441 atcaccgaaa cgcgcgaggc agcagatcaa ttcgcgcgcg aaggcgaagc ggcatgcatt
     4501 tacgttgaca ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa
     4561 gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat
     4621 gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg
     4681 aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg
     4741 gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc
     4801 ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc
     4861 agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac
     4921 aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat
     4981 gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac
     5041 cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag
     5101 catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc
     5161 tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg
     5221 atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg
     5281 ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc
     5341 gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga
     5401 tacgacgata ccgaagacag ctcatgttat atcccgccgt caaccaccat caaacaggat
     5461 tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg
     5521 gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc
     5581 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     5641 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agcgcgaatt
     5701 gatctg
//
