LOCUS       sequence-483456-        5442 bp    DNA     circular SYN 15-AUG-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5442)
  AUTHORS   Liau SS, Jazag A, Whang EE
  TITLE     HMGA1 is a determinant of cellular invasiveness and in vivo 
            metastatic potential in pancreatic adenocarcinoma.
  JOURNAL   Cancer Res. 2006 Dec 15. 66(24):11613-22.
  PUBMED    17178855
REFERENCE   2  (bases 1 to 5442)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cancer Res.
            2006 Dec 15. 66(24):11613-22."
COMMENT     Sequence Label: pIRES-HMGA1
FEATURES             Location/Qualifiers
     source          1..5442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          1088..1317
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     1300..1319
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(1390..1964,1965..1967,1968..1976)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1557..1574)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1784..1803
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1996..2595
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1996..2015)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2492..2512
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    2829..2950
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2866..2885)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3051..3067)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3051..3067)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3064..3086)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3075..3091
                     /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)."
     promoter        complement(join(3099..3105,3106..3123,3124..3129))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3144..3165
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3282..3299)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3453..4041)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3533..3552)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4212..5003,5004..5072))
                     /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     4835..4854
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5073..5177)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5252..5271)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatcgat atcatgagtg agtcgagctc gaagtccagc cagcccttgg
      721 cctccaagca ggaaaaggac ggcactgaga agcggggccg gggcaggccg cgcaagcagc
      781 ctccggtgag tcccgggaca gcgctggtag ggagtcagaa ggagcccagc gaagtgccaa
      841 cacctaagag acctcggggc cgaccaaagg gaagcaaaaa caagggtgct gccaagaccc
      901 ggaaaaccac cacaactcca ggaaggaaac caaggggcag acccaaaaaa ctggagaagg
      961 aggaagagga gggcatctcg caggagtcct cggaggagga gcagtgagaa ttcggatccg
     1021 cggccgcata gataactgat ccagtgtgct ggaattaatt cgctgtctgc gagggccagc
     1081 tgttggggtg agtactccct ctcaaaagcg ggcatgactt ctgcgctaag attgtcagtt
     1141 tccaaaaacg aggaggattt gatattcacc tggcccgcgg tgatgccttt gagggtggcc
     1201 gcgtccatct ggtcagaaaa gacaatcttt ttgttgtcaa gcttgaggtg tggcaggctt
     1261 gagatctggc catacacttg agtgacaatg acatccactt tgcctttctc tccacaggtg
     1321 tccactccca ggtccaactg caggtcgagc atgcatctag ggcggccaat tccgcccctc
     1381 tccccccccc ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata
     1441 aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt
     1501 gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct
     1561 cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc
     1621 ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga
     1681 caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc
     1741 ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt
     1801 attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg
     1861 gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc
     1921 gaaccacggg gacgtggttt tcctttgaaa aacacgatga taagcttgcc acaacccaca
     1981 aggagacgac cttccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     2041 gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     2101 accgtcgacc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     2161 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     2221 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     2281 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
     2341 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     2401 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
     2461 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     2521 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     2581 aagcccggtg cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga
     2641 ccccatggct ccgaccgaag ccacccgggg cggccccgcc gaccccgcac ccgcccccga
     2701 ggcccaccga ctctagataa ctgatcataa tcagccatac cacatttgta gaggttttac
     2761 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     2821 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     2881 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     2941 atgtatctta acgctcgagt gcattctagt tgtggtttgt ccaaactcat caatgtatct
     3001 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     3061 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     3121 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     3181 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     3241 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     3301 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3361 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3421 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3481 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3541 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3601 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3661 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3721 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3781 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3841 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3901 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3961 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4021 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4081 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4141 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4201 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4261 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4321 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4381 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4441 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4501 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4561 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4621 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4681 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4741 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4801 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4861 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4921 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4981 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5041 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5101 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5161 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     5221 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     5281 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
     5341 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
     5401 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg tt
//
