LOCUS       Exported                5063 bp DNA     circular SYN 21-JUL-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    pHL-sec
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5063)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5063)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5063
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      31..40
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     37..129
                     /codon_start=1
                     /label=Sig_pep
                     /translation="MGILPSPGMPALLSLVSLLSVLLMGCVAETG"
     CDS             130..561
                     /codon_start=1
                     /product="Human ephrin b2 (EFNB2)
                     "
                     /label=Ephrin RBD
                     /translation="SIVLEPIYWNSSNSKFLPGQGLVLYPQIGDKLDIICPKVDSKTVG
                     QYEYYKVYMVDKDQADRCTIKKENTPLLNCAKPDQDIKFTIKFQEFSPNLWGLEFQKNK
                     DYYIISTSNGSLEGLDNQEGGVCQTRAMKILMKVGQDGTK"
     CDS             562..579
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    756..811
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1172..1188)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1196..1212
                     /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(1220..1250)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1265..1286
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1582..2170)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2341..3201)
                     /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"
     promoter        complement(3202..3306)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3337..3716
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3718..3995
                     /label=chicken beta-actin promoter
     intron          3996..5012
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
ORIGIN
        1 ttgcggccgt ctcaggccga attcaagctt gccaccatgg ggatccttcc cagccctggg
       61 atgcctgcgc tgctctccct cgtgagcctt ctctccgtgc tgctgatggg ttgcgtagct
      121 gaaaccggtt cgatagtttt agagcctatc tattggaatt cctcgaactc caaatttcta
      181 cctggacaag gactggtact atacccacag ataggagaca aattggatat tatttgcccc
      241 aaagtggact ctaaaactgt tggccagtat gaatattata aagtttatat ggttgataaa
      301 gaccaagcag acagatgcac tattaagaag gaaaataccc ctctcctcaa ctgtgccaaa
      361 ccagaccaag atatcaaatt caccatcaag tttcaagaat tcagccctaa cctctggggt
      421 ctagaatttc agaagaacaa agattattac attatatcta catcaaatgg gtctttggag
      481 ggcctggata accaggaggg aggggtgtgc cagacaagag ccatgaagat cctcatgaaa
      541 gttggacaag atggtaccaa gcaccaccat caccatcact aatgatcact cgagactagt
      601 atcgcgataa ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc
      661 caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga
      721 catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc
      781 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
      841 tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc
      901 tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt
      961 ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt
     1021 tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac tagccagatt
     1081 tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct
     1141 cgacctgcag cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     1201 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     1261 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     1321 gaaacctgtc gtgccagcgg atccgctgca ttaatgaatc ggccaacgcg cggggagagg
     1381 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     1441 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     1501 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     1561 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     1621 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     1681 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     1741 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     1801 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     1861 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     1921 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     1981 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     2041 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     2101 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2161 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     2221 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     2281 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     2341 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     2401 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     2461 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     2521 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     2581 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     2641 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     2701 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     2761 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     2821 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     2881 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     2941 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     3001 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     3061 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     3121 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     3181 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     3241 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     3301 tccgcgcaca tttccccgaa aagtgccacc tgggtcgaca ttgattattg actagttatt
     3361 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     3421 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     3481 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     3541 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     3601 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     3661 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg
     3721 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt
     3781 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggggc
     3841 gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg
     3901 cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc
     3961 ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc
     4021 ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac
     4081 tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt
     4141 aatgacggct tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc
     4201 ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg
     4261 cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg
     4321 cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg
     4381 gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg
     4441 tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc tccccgagtt gctgagcacg
     4501 gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg
     4561 gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc ggggagggct
     4621 cgggggaggg gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca
     4681 gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct
     4741 gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc
     4801 ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc
     4861 gtccccttct ccctctccag cctcggggct gtccgcgggg ggacggctgc cttcgggggg
     4921 gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta
     4981 accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc
     5041 tgtctcatca ttttggcaaa gaa
//
