LOCUS       sequence-6858-ex        4988 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 4988)
  AUTHORS   Shoichet SA, Malik TH, Rothman JH, Shivdasani RA
  TITLE     Action of the Caenorhabditis elegans GATA factor END-1 in Xenopus 
            suggests that similar mechanisms initiate endoderm development in 
            ecdysozoa and vertebrates.
  JOURNAL   Proc Natl Acad Sci U S A. 2000 Apr 11. 97(8):4076-81.
  PUBMED    10760276
REFERENCE   2  (bases 1 to 4988)
  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: "Proc Natl 
            Acad Sci U S A. 2000 Apr 11. 97(8):4076-81."
COMMENT     Sequence Label: p026 pCS2-EnR
FEATURES             Location/Qualifiers
     source          1..4988
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    941..1075
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(965..984)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1019..1038
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1192..1212)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1192..1210)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1231..1247)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1231..1247)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1244..1266)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1255..1271
                     /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(1279..1285,1286..1303,1304..1309))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1324..1345
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1462..1479)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1633..2221)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1713..1732)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2392..3183,3184..3252))
                     /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     3015..3034
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3253..3357)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3383..3838)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3520..3541)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3732..3751
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3910..4894
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        4929..4947
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4929..4946
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 aattcagatc tctcgaggcc ctggaggatc gctgcagccc acagtcagcg cccagcccca
       61 ttaccctaca aatgcagcat cttcaccacc agcaacagca gcagcagcaa cagcagcagc
      121 aaatgcagca cctccaccag ctgcagcaac tgcagcagtt gcaccaacag caactggccg
      181 ccggtgtctt ccaccatccg gcaatggcct tcgatgccgc tgcagccgcc gctgctgcag
      241 ctgctgctgc ggccgcccac gctcatgctg ctgcactgca gcagcgcctc agtggcagtg
      301 gatcgcccgc atcctgctcc acgcccgcct cgtccacgcc gctgaccatc aaggaggagg
      361 aaagcgactc cgtgatcggt gacatgagtt tccacaatca gacgcacacc accaacgagg
      421 aggaggaggc ggaggaggat gacgacattg atgtggatgt ggatgatacg tcggcgggcg
      481 gacgcctgcc accacccgcc caccagcagc agtcgacggc caagccctcg ctggcctttt
      541 ccatctccaa catcctgagc gatcgtttcg gagatgtcca gaagcctggc aagtcgattg
      601 agaaccaggc cagcatattc cgccccttcg aggcgaatcg ctcccagact gccacgccct
      661 ccgcctttac aagagtggat ctgctggagt ttagccggca acagcaagct gccgccgcag
      721 ccgctactgc ggccatgatg ctggaacggg ccaacttcct taactgcttc aatccggctg
      781 cctatcccag gatacacgag gaaatcgtgc agagtcgcct gcgcaggagt gcagccaatg
      841 ccgtcatccc gccgcccatg agctccaaga tgagcgatgc caatccagag aaatctgctc
      901 tgggatctta atctagaact atagtgagtc gtattacgta gatccagaca tgataagata
      961 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     1021 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     1081 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaatt
     1141 cgcggcgcgc cgcggcgcca atgcattggg cccggtaccc agcttttgtt ccctttagtg
     1201 agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     1261 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     1321 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     1381 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     1441 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     1501 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     1561 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     1621 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1681 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1741 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1801 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     1861 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     1921 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     1981 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2041 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     2101 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2161 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2221 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     2281 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     2341 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     2401 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     2461 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     2521 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     2581 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     2641 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     2701 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     2761 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     2821 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     2881 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     2941 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     3001 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     3061 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     3121 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     3181 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     3241 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     3301 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     3361 atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta
     3421 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     3481 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     3541 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     3601 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     3661 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     3721 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     3781 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
     3841 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     3901 ttacgccagg accatagcca attcaatatg gcgtatatgg actcatgcca attcaatatg
     3961 gtggatctgg acctgtgcca attcaatatg gcgtatatgg actcgtgcca attcaatatg
     4021 gtggatctgg accccagcca attcaatatg gcggacttgg caccatgcca attcaatatg
     4081 gcggacctgg cactgtgcca actggggagg ggtctacttg gcacggtgcc aagtttgagg
     4141 aggggtcttg gccctgtgcc aagtccgcca tattgaattg gcatggtgcc aataatggcg
     4201 gccatattgg ctatatgcca ggatcaatat ataggcaata tccaatatgg ccctatgcca
     4261 atatggctat tggccaggtt caatactatg tattggccct atgccatata gtattccata
     4321 tatgggtttt cctattgacg tagatagccc ctcccaatgg gcggtcccat ataccatata
     4381 tggggcttcc taataccgcc catagccact cccccattga cgtcaatggt ctctatatat
     4441 ggtctttcct attgacgtca tatgggcggt cctattgacg tatatggcgc ctcccccatt
     4501 gacgtcaatt acggtaaatg gcccgcctgg ctcaatgccc attgacgtca ataggaccac
     4561 ccaccattga cgtcaatggg atggctcatt gcccattcat atccgttctc acgcccccta
     4621 ttgacgtcaa tgacggtaaa tggcccactt ggcagtacat caatatctat taatagtaac
     4681 ttggcaagta cattactatt ggaagtacgc cagggtacat tggcagtact cccattgacg
     4741 tcaatggcgg taaatggccc gcgatggctg ccaagtacat ccccattgac gtcaatgggg
     4801 aggggcaatg acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc gtgcctaatg
     4861 ggaggtctat ataagcaatg ctcgtttagg gaaccgccat tctgcctggg gacgtcggag
     4921 caagcttgat ttaggtgaca ctatagaata caagctactt gttctttttg caggatccca
     4981 tcgattcg
//
